Cholesterol Homeostasis

Home , Brassicasterol, Esterase, Mevalonic acid, Stigmastanol

FOR LIFE SCIENCE RESEARCH Volume 2 Number 7

■ HMGR Assay Kit

■ Cholesterol Biosynthesis

■ Blocking Absorption of Dietary Cholesterol Hypercholesterolemia can lead to ■ Cholesterol Esterification the formation of plaques and the development of atherosclerosis. ■ Cholesterol Transport

■ Bile Acids Lipid Resource

Sigma has now concentrated all its lipids and lipid related products into one easy-to-navigate location. Quickly find the specific lipids you need from over 2,000:

■ Fatty Acids ■ Sphingolipids ■ Glycerides ■ Prenols ■ Complex Lipids ■ Fluorescent Labeled Lipids ■ Oils ■ Analytical Standards ■ Bioactive Lipids

Browse for lipids in cell signaling using our interactive pathway charts. Discover everything researchers need for lipid research at sigma.com/lipids.

sigma-aldrich.com 1

Introduction Cholesterol is an essential biological molecule that performs many functions within the body. It is a structural component of all cell membranes and is also a precursor to steroid hormones, vitamin D, and bile acids that aid in digestion. Within membranes FOR LIFE SCIENCE RESEARCH the cholesterol to polar lipid ratios affect stability, permeability, and protein mobility. The hormones produced from cholesterol include androgens, estrogens, and the gluco- and 2007 mineralocorticoids. Volume 2 Cholesterol levels in the body are achieved via two sources. Adults with healthy diets will biosynthesize the majority of their cholesterol in the liver and other body tissues Number 7 and obtain the remainder from the dietary intake of foods high in saturated fatty acids. Free cholesterol is not found in blood; rather it is esterified to fatty acids and packaged in lipoprotein particles. Very low density lipoproteins (VLDL) are produced by Table of Contents the liver and consist of an outer core composed of apolipoproteins; apo-B100, apo-CI, apo-CII, apo-CIII, and apoE surrounding an inner core of phospholipids, triglycerides, cholesterol, and cholesteryl esters. In the blood, VLDL transfers apolipoprotein-CII to Cholesterol Homeostasis high density lipoprotein (HDL) and lipoprotein lipase in the capillaries begins to remove the triglycerides, transforming the particle into an intermediate density lipoprotein (IDL). About 50% of IDL particles are removed from the circulation by the liver. The remaining IDLs are transformed to low density lipoprotein particles (LDL, the so-called “bad” cholesterol) by the loss of apolipoprotein E and the further reduction of triglyceride Introduction ...... 1 content until it is exceeded by the content of cholesteryl esters. LDL particles deliver lipids to the body’s cells via LDL receptor-mediated endocytosis. When LDL lipids are HMG-CoA Reductase (HMGR) Assay Kit ...... 2 oxidized by free radicals, they bind more easily to the proteoglycans lining the vascular endothelium, and thus, become incorporated into atherosclerotic plaque. Cholesterol Biosynthesis ...... 3 HDL, the so-called “good” cholesterol is composed of apolipoproteins-CII and E Biosynthesis Regulation ...... 6 surrounding a lipid core. HDL particles circulate through the capillaries collecting lipids Statins ...... 8 including cholesterol and cholesteryl esters and returning them to the liver for further metabolism. Cholesterol returned to the liver by HDL is synthesized into bile acids. Bile Blocking Absorption of Dietary acids facilitate the digestion of lipids by acting as emulsifying agents and also aid in the Cholesterol ...... 9 absorption of fat-soluble vitamins. Cholesterol is ultimately excreted from the body as bile acids. Cholesterol Esterification ...... 10 Excessive levels of oxidized LDL in the blood can lead to potential health risks. Normally Cholesterol Esterase ...... 12 cholesterol levels are tightly controlled by complex mechanisms. When dietary intake of cholesterol is high, biosynthesis is reduced. However, the body’s homeostatic Cholesterol Transport ...... 13 mechanisms can be inadequate when baseline endogenous cholesterol biosynthesis Lipoprotein Regulation ...... 16 becomes excessive or when dietary cholesterol intake is overwhelming. For these instances, drugs have been discovered that can reduce cholesterol biosynthesis (statins), Bile Acids ...... 17 reduce the intestinal absorption of dietary cholesterol and other lipids (ezetimibe), or enhance the metabolic utilization of lipids in the liver (fibrates). These drugs serve to keep the blood levels of LDL in check to avoid the deleterious effects that can arise from the accumulation of vascular plaque, including such serious medical conditions as atherosclerosis, coronary artery disease, and stroke. This issue of BioFiles highlights the product groups that Sigma offers to further the research of cholesterol absorption, biosynthesis, transport, and excretion. We introduce two key research tools in the HMG-CoA Reductase enzyme and assay kit. We also showcase a number of important cholesterol lowering molecules including statins, sterols, and stanols. HMG-CoA Reductase 2 sigma.com HMG-CoA Reductase(HMGR)AssayKit contributes tothedevelopmentofachronic inflammatorystate. evidence suggeststhatadisturbanceofcholesterol homeostasis which mightresult inmyocardial infarction andstroke. Recent atherosclerosis andconsequentlytocardiovascular pathologies, role ashypercholesterolemia oftenleadstothedevelopmentof Controlling serumcholesterol levelshasanimportanttherapeutic Figure 1. Applications HMG-CoA. by thecatalyticsubunitofHMGRinpresence ofthesubstrate decrease inabsorbance,whichrepresents theoxidationofNADPH The assayisbasedonaspectrophotometric measurement ofthe HMG-CoA +2NADPH2H Principle oftheAssay Kit Features andBenefits reticulum. transmembrane glycoprotein, locatedontheendoplasmic 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR)isa in theclinicaltreatment ofhighserumcholesterol levels. the humanenzymehasbeentargetedsuccessfullybydrugs products. Inadditiontothephysiologicalregulation ofHMGR, in order tomaintaintheconcentrationofmevalonatederived controlled through synthesis,degradation,andphosphorylation limiting stepinsterol biosynthesis. HMG-CoA tocoenzymeAandmevalonate,whichistherate-

Absorption 340 nm ■ ■ Reaction scheme: ■ ■ ■ ■ Sufficient for30assaysof1mLor100200µL Includesacontrol inhibitor 0.2 0.4 0.6 0.8 1.2 1.4 1.6 role intherapeutics) inhibitors/activators oftheenzyme(whichmayplayacrucial Detection ofHMGRactivityaswellscreening fordifferent pathways Basic research ofcholesterol andotherrelated metabolic HMGR inhibitorsandactivators Includes anHMGRcontrol enzyme,enablingscreening of of HMGRenzymeactivity Contains allthereagents forasimpleandrapidmeasurement 0 1 InhibitionofHMG-CoAreductase (HMGR)activitybyPravastatin. 01 21 415 14 13 12 11 10 9 8 7 6 5 4 3 2 1 0 1 Thisenzymecatalyzesthefour-electron reduction of w/o inhibitor w/o inhibitor 5nMPravastatin 25 nMPravastatin 50 nMPravastatin 250 +

➞ HMGR

mevalonate+2NADP 2 Time (min) TheactivityofHMGRis + 3,4 +CoASH

2. 1. References protocol. 96 wellplate,according totheHMG-CoAReductaseAssayKit performed inaUVcompatible is aspecificinhibitorofHMG-CoAreductase. Theassaywas HMG-CoA, anddifferent concentrationsofPravastatin,which was incubatedat37°Cwith400µMNADPH,0.3mg/ml Approximately 6µgofHMGRenzyme(CatalogNumberH7039) 3. 5. 4. Kit Components Kit Components CS1090 Inhibitor Solution(Pravastatin) (0.55-0.65 mg/mL) HMG-CoA Reductase(catalytic domain) Substrate Solution(HMG-CoA) NADPH Cat.No. Assay Buffer, 5 a.N.PoutNm PackSize PackSize Product Name Cat. No. Product Name rosuvastatin. 3-hydroxy-3-methylglutaryl CoA(HMG-CoA)reductase by Holdgate, G.A.,et.al.,Molecularmechanismforinhibitionof membrane proliferations. elevated levelscorresponds todistinctendoplasmicreticulum Saccharomyces cerevisiae Koning, A.J.,et.al.,Different subcellularlocalizationof and catalysis. human HMG-CoAreductase: insightsintoregulation ofactivity Istvan, E.S.,et.al.,Crystalstructure ofthecatalyticportion Targets Cardiovas.Haemat.,Disord. atherosclerosis inducedbydietarycholesterol. effects onchronic subacuteinflammationandonsetof Kleemann, R.,andKooistra,T., HMG-CoAreductase inhibitors: 1529 portion ofhumanHMG-CoAreductase. Istvan, E.S.,andDeisenhofer, J.,Thestructure ofthecatalytic , 9-18(2000). M-o euts sa i 1kit HMG-CoA ReductaseAssaykit Biochem. Soc.Trans. 3 EMBO J., N6505

19 HMG-CoAreductase isozymesat Mol. Biol , 819-830(2000). , . Cell, 31 , 5 , 528-531(2003). , 441-453(2005). Biochim. Biophys.Acta 7 , 769-789(1996). Curr. Drug 200 µL 200 µL 25 mg 10 mL 2 mL , Cholesterol Biosynthesis 3

sigma.com . sigma.com/metapath HMG-CoA is then converted to mevalonate by HMG-CoA by HMG-CoA then converted to mevalonate HMG-CoA is with the aid of is completed This reaction (HMGR). reductase reactions for all reduction is used as a cofactor NADPH, which a series undergoes synthesis. Mevalonate cholesterol throughout yielding the isoprenoid, and a decarboxylation of phosphorylations reactions (IPP). A series of condensing isopentenyl pyrophosphate by squalene synthase, leading to the production catalyzed occur, is the first of the sterols squalene, lanosterol, of squalene. From 19 requires to cholesterol of lanosterol formed. The conversion steps. additional reaction IUBMB-Sigma-Nicholson Metabolic Pathways Chart. This chart can be viewed in detail at Figure 1. Figure

intake and biosynthesis. The majority of cholesterol utilized by utilized by of cholesterol The majority intake and biosynthesis. ~70% of which produces liver, is synthesized in the healthy adults The other 30% (~1 gram). requirement cholesterol the total daily intake. dietary comes from generally takes place in the Biosynthesis of cholesterol cells and begins with acetyl- of hepatic endoplasmic reticulum in the an oxidation reaction from CoA, which is mainly derived also be derived from acetyl-CoA can mitochondria. However, of ethanol by acetyl-CoA synthetase. the cytoplasmic oxidation converted to 3-hydroxy- are Acetyl-CoA and acetoacetyl-CoA by HMG-CoA synthase. 3-methylglutaryl-CoA (HMG-CoA)

Cholesterol levels in the body come from two sources, dietary sources, two in the body come from levels Cholesterol Cholesterol Biosynthesis Cholesterol Cholesterol Biosynthesis 4 sigma.com  Supplied asasolutioncontaining250µgprotein in50mMTris Key intermediateinthebiosynthesisofterpenes,cholesterol, and 3-Hydroxy-3-methyl The conversionofHMG-CoAtomevalonateby One unitwillconvert1.0µmoleofNADPHtoNADP activity: 2-8 units/mg protein store at: store at: ketone bodies. Consists ofasinglepolypeptidechain888aminoacids.The Hydroxy-3-methylglutaryl-CoA reductase catalyzesthecommitted ≥ (Catalog NumberP8340)and50%(w/v)glycerol. pH 7.5,with5mMDTT, 1:200Protease InhibitorCocktail and isunderstrictregulatory control (see reductase istherate-limitingstepofcholesterol biosynthesis at 37°C. connects thetwoportionsofprotein. in itscytoplasmic,solublecarboxy-terminalportion.Alinkerregion reticulum membrane,whilethecatalyticsiteofprotein resides residues are membraneboundandreside intheendoplasmic single polypeptidechainof888aminoacids.Theamino-terminal cholesterol levels.HumanHMG-CoAreductase consistsofa the targetofcompoundsthatare effective inloweringserum DL linker region connectsthetwoportionsofprotein. protein resides initscytoplasmic,solubleC-terminal portion.A endoplasmic reticulum membrane,whilethecatalytic siteofthe amino-terminal residues are membraneboundandreside inthe step incholesterol biosynthesis. Figure 1. C HMG-CoA HMG-CoA Reductase 27 90% (SDS-PAGE) H6132-25MG H6132-10MG H6132-5MG H7039-250UG min. 90%

-3-Hydroxy-3-methyl

H 44 4080120160200240280320360400440480520560600640680720760800840880 Region Transmembrane

N 7 B A

O HMG-CoAReductase. 20

P 3

Transmembrane Region (Human Sequence) FW 1009.67 ; HMGR glutar glutar

Linker

yl-CoA reductase human [103476-21-7] yl coenzyme A sodium salt coenzyme Asodium yl Catalytic Domain Figure 1 Domain Catalytic

). HMGRis + perminute

Linker

250 µg 8 25 mg 10 mg 5 mg     Iso (±)-Mevalono Geranyl pyro

store at: store at: store at: store at: vial of200 µg Intermediate interpenebiosynthesis vial of200 µg Intermediate interpenebiosynthesis vial of200 µg Intermediate interpenebiosynthesis.

γ ≥ ≥ ≥ lactone

trans

C IPP (±)- C DMAPP C C , 5 6 10 5 95% (TLC) 90% (TLC) 95% (TLC) I0503-5VL I0503-1VL M4667-10G M4667-5G M4667-1G G6772-5VL G6772-1VL D4287-5VL D4287-1VL γ 1 mg/mL inmethanol:aqueous10mMNH ~97% (titration) 1 mg/mL inmethanol:aqueous 10mMNH 1 mg/mL inmethanol:aqueous 10mMNH

H H H

H β penten -Dimethyl 12 10 12 20 -Hydroxy-

-3,7-Dimethyl-2,6-octa

O O O

; 7 3 7

7 P P

B B B B

P 2 2

2 ·3NH ·3NH FW 130.14

-Mevalo

·3NH yl pyro β

allyl pyro -methyl- phos 3 3 lactone

lactone FW 297.18 FW 297.18 phos

phate ammonium salt phate ammonium FW 365.30 [674-26-0] δ -valero

; phate triammonium salt solution phate triammonium DL

phos dienyl pyro

-Mevalonic acidlactone

lactone [116057-53-5] [1186-30-7]

phate triammonium salt phate triammonium

[763-10-0] phos

;

(±)-3-Hydroxy-3-methyl phate

;

GPP 4 4 4 OH (7:3), OH (7:3), OH (7:3),

δ -valero- 5 vials 5 vials 5 vials 1 vial 1 vial 1 vial 10 g 5 g 1 g Cholesterol Biosynthesis 5

sigma.com 1 g 5 g 1 g 5 g 1 kg 5 kg 10 g 25 g 25 g 25 g 100 g 500 g 100 g 100 g 500 g 50 mg 500 mg 500 mg tene 99% es ≥

[57-88-5] -Hydroxy-5-chol -Hydroxy-5-chol

β 95% (GC), Ash free 95% (GC), Ash free 3

≥ ;

98% -ol

≥

® β ol FW 386.65 l o

B E E ten-3 ter ter es ter ter O

es 46 es H

from bovine, Sigma Grade, bovine, Sigma Grade, from synthetic, from bovine, bovine, from Grade I, ~99% liver, porcine from sheep wool, ~95% (GC) from C8503-5KG C8667-1G C9913-500MG C8503-100G C8503-500G C8503-1KG C8667-500MG C8667-5G C8667-25G C9913-50MG C3292-10G C3137-1G C3137-5G C3137-25G C8503-25G C8667-100G C3292-100G C3292-500G 27

Chol 5-Chol C

brain lipid is cholesterol.

total membranes; ~25% of of all biological Major component store at: store at: store store at: store

Precipitated from alcohol from Precipitated chromatography for Standard Formerly Cat. No. CH-PL Equivalent to USP/NF Synthesized from material of non-animal origin material of non-animal origin Synthesized from SyntheChol Chol      1 L es- 1 vial 1 mg 5 mg 5 mg

5 vials 10 mL 10 mg 100 mL 500 mL ene [111-02-4]

OH (7:3),

hexa 4 phate ammonium phate ammonium -Hydroxy-5,24-chol -Hydroxy-5,24-chol -ol cosa β β phos 3

;

ol FW 410.72

ter ter es

2 -]

chol 2

trien-1-yl pyro trien-1-yl pyro

8,24-Lanostadien-3

;

=CHCH

2 [13058-04-3] )]

24-Dehydro 24-Dehydro

diene ;

[313-04-2] [79-63-0] phate ammonium phate ammonium salt

-ol methyl-2,6,10,14,18,22-tetra C(CH

2 β CH

phos 2 FW 433.42

methyl-2,6,10-dodeca methyl-2,6,10-dodeca

2 tadien-3 P

FW 384.64 FW 426.72 7 es O

B B E B 3

C[=CHCH

O O N

2 )

44 50 37 3,7,11-Tri 3,7,11-Tri

3 85% (GC) 98% ;

H H H

-Hydroxy-8,24-lanosta -Hydroxy-8,24-lanosta ≥

≥

from sheep wool, ~97% from Solution in methanol: 10 mM aqueous NH Solution in methanol: D6513-10MG D6513-5MG L5768-1MG L5768-5MG S3626-10ML S3626-100ML F6892-1VL F6892-5VL S3626-500ML S3626-1L diene 95% (TLC) 27 30 15 β C salt C 5,24-Chol C 3 2,6,10,15,19,23-Hexa FPP

[(CH

Actual concentration given on label Actual concentration given including mass G proteins, of several low molecular prenylation Ras. ≥

and antitumor agents. cells exposed to carcinogens

vial of 200 µg vial of 200 Isoprenoid from the intracellular mevalonate pathway used for the intracellular mevalonate pathway from Isoprenoid store at: store store at: store store at: store store at: store

sterol precursor Cholesterol

to normal Cytoprotective to all steroids. Biosynthetic precursor

Desmosterol Desmosterol

Lanosterol Lanosterol

Squalene

Farnesyl pyro     Cholesterol Biosynthesis 6 sigma.com 

store at: AY 9944

Cholesterol synthesis inhibitor;inhibitsboth7-dehydrocholesterol The amountofcholesterol thatissynthesizedinthelivertightly Δ trans clearing evenmore cholesterol from theplasma. in cholesterol synthesisalsopromotes anincrease ofHDL,thus, plasma LDLcholesterol anditsrelated healthrisks.Thedecrease the clearancerateofLDLparticlesfrom theplasmaandreduces the densityofLDLreceptors ontheirsurfaces.Thisincreases synthesis leadstoahomeostaticresponse inwhichcellsincrease amount ofcholesterol produced bytheliver. Limitingcholesterol regulation ofkeyenzymessuchasHMGR, thus,reducing the ultimately stopthematurationofSREBPs,resulting inthedown CoA reductase (HMGR). Highdietarysterol levelsactingonSCAP biosynthetic pathwayincludingtheratelimitingenzymeHMG- SREBPs servetoregulate all12enzymesinthecholesterol promoting geneexpression. complex remains intheERpreventing cleavedSREBPfrom gene expression. Whenoxysterol levelsare hightheSCAP/SREBP move intothenucleus,where itinteractswithSREstopromote to theGolgiwhere SREBPiscleavedandaportionofitcannow When oxysterol levelsare lowtheSCAP/SREBPcomplexmoves SCAP responds tothelevelofoxysterols present inthecell. the endoplasmicreticulum (ER)where aregulatory domainwithin cleavage activatingproteins (SCAPs).SCAPsbindtoSREBPsin regulating proteins are boundbyanotherprotein calledSREBP instrumental role incholesterol homeostasis.Thesetranscription Proteins (SREBPs).DuetotheirabilitybindSREs,SREBPsplayan important toactivatingSREsare Sterol RegulatingElementBinding biosynthetic pathwayandLDLreceptors. Transcription factors promoters ofthegenescodingforenzymescholesterol Sterol-Sensitive ResponseElements(SREs).SREsare foundinthe enzymes inthecholesterol biosyntheticpathwayisdependenton LDL receptor expression andcontrolling theexpression ofallthe density lipoprotein (LDL)particles.Onemechanismforregulating LDL receptors regulate thecellulartransportoflipidrichlow levels ofcholesterol. similar oxysterols actasregulatory moleculestomaintainhealthy other tissuesisundernosuchfeedbackcontrol. Cholesterol and is low, synthesisisincreased. However, cholesterol produced in cholesterol ishigh,synthesisdecreased andwhendietaryintake regulated bydietarycholesterol levels.Whendietaryintakeof Biosynthesis Regulation

C solubility 22 7-reductase and C5364-25MG C5364-5MG ≥

H 99% (HPLC) 30

-1,4-bis(2-Chloro

Cl 4 E

DMSO...... 14mg/mL

N 2

FW 464.30

Δ benzyl 8,7-sterol isomerase.

[366-93-8] amino ethyl) cyclo

hexane dihydrochloride 25 mg 5 mg    Brassicasterol BM 15766 sulfate Antihyperlipoproteinemic believedtoactbyinhibitingcholesterol Clofibrate Cerulenin

store at: store at: store at: store at: Brassicasterol isaphytosterol found inrapeseedandcanolaoils; Dehydrocholesterol reductase inhibitor. Antibiotic; fattyacidsynthaseinhibitor. biosynthesis. the mammaliancholesterol biosynthesispathway.1 been showntoinhibitsterol it isalsopresent inmarinealgaeandshellfish.Brassicasterol has last stepofcholesterol synthesis. Inhibits 7-dehydrocholesterol acid sulfate

phen

5,22-Chol C 4-[2-[4-[3-(4-Chloro 2-(4-Chloro (2 C C C 28 22 12 12 R B8685-5MG B8685-1MG C6643-10G C6643-5G C6643-1G C6643-250MG C2389-50MG C2389-10MG C2389-5MG B4936-5MG from semisynthetic ~95%, from ≥

H H H H

,3 98% (HPLC) 46 25 15 17 oxy)iso S

O ClN ClO NO

, E

, E E E B

E 3 es 2 3

)-2,3-Epoxy-4-oxo-7,10-dodeca

O FW 398.66

phen butyrate tadien-24 2 FW 223.27 FW 242.70

·H

oxy)-2-methyl 2 Cephalosporium caerulens

SO phenyl)-2-prop

; 4

Ethyl 2-(4-chloro β

-methyl-3

FW 482.98 [474-67-9]

[17397-89-6] [637-07-0] pro β Δ pionic acidethylester Δ enyl]-1-piperazinyl]ethyl]benzoic -ol 24

7-reductase, whichcatalyzesthe

[86621-94-5] -reductase, anenzymeinvolvedin phen

dien oxy)-2-methyl amide

;

Ethyl 2-(4-chloro- pro pion ate 250 mg 50 mg 10 mg 5 mg 5 mg 1 mg 5 mg 10 g 5 g 1 g Cholesterol Biosynthesis 7

sigma.com nate 1 mg 5 mg 5 mg 5 mg 25 mg 25 mg pho one 24- phos Δ phen

benzo 60 °C ≤

phenyl)ethenyl-1,1-bis [189197-69-1] st-5-en-17-one dihydrochloride st-5-en-17-one dihydrochloride

[3039-71-2]

10 mg/mL sigma.com/biofiles oxy]-4-bromo-2’-fluoro oxy]-4-bromo-2’-fluoro ≥ [126411-39-0] FW 564.44

-butyl-4-hydroxy -butyl-4-hydroxy

Visit Visit 4 O

FW 460.52 4 tert

amino)hexyl 4 amino)ethoxy]andro amino)ethoxy]andro

FW 504.53 O...... insoluble O...... 9 mg/mL at O...... >5 mg/mL O...... >5 at ~60 °C

2 2 2

methyl · 2HCl BrF · C

2 2 2 H H H

E E 7 NO NO O

41 27 42 ethyl 2-(3,5-di- 98% (HPLC) 98% )-3-[2-(Diethyl H H H

≥ ≥ S4194-5MG S4194-1MG U3633-5MG U3633-25MG R2278-5MG R2278-25MG β 25 23 24 solubility DMSO......

solubility solubility C C (3 [4’-[6-(Allyl Tetra C

reductase); Weak inhibitor of hedgehog (hh) signaling. inhibitor of hedgehog Weak reductase);

a cyclase (OSC, E.C. 5.4.99.7) represents Oxidosqualene:lanosterol Partial inhibition of lowering drug. unique target for a cholesterol sterols, and subsequent synthesis of lanosterol OSC should reduce that repress of epoxysterols production and also stimulate the a synergistic, self- generating expression, HMG-CoA reductase loop. limited negative regulatory fumarate (1:1) fumarate (1:1) synthesis (inhibits desmosterol Inhibitor of cholesterol 2,3- cyclase inhibitor. Orally active 2,3-oxidosqualene:lanosterol inhibitor. lowering drug; HMGCoA reductase Cholesterol

store at: store store at: store U 18666A Ro 48-8071 fumarate Ro 48-8071 SR 12813   Read current and previous issues and register to issues and register and previous Read current BioFiles issues. future receive Your gateway to products, services and more services and more gateway to products, Your Life Science Research ta- 1 g 5 g es 25 g 1 mg 5 mg 10 mg 25 mg 10 mg 100 mg conta-2,6,8,10,12, -Hydroxy-5,7-chol -Hydroxy-5,7-chol β 3

;

ol ter ter methyldotria es chol

ene deca

ol (−)-7-Dehydro (−)-7-Dehydro

ol [2140-46-7] [434-16-2] ;

A ter ter

[502-65-8] ter ter -ol

es 3 β es ,25-diol β 2,6,10,14,19,23,27,31-Octa

tore at: tore

chol ;

chol FW 402.65 FW 384.64 tadien-3

B tene-3 FW 536.87 es 2 Provitamin D Provitamin

O O

pene ;

56 46 44 90%, from tomato 90%, from 98% 85% -Carotene -Carotene H H H

ψ ≥ ≥ ≥ H1015-25MG H1015-100MG L9879-1MG L9879-5MG L9879-10MG H1015-10MG D4429-1G D4429-5G D4429-25G , 40 27 27 C C 5-Chol 5,7-Chol C 14,16,18,20,22,24,26,30-tri

ship: dry ice s

fibroblasts. skin Opitz syndrome

diene

induces apoptosis (SREBPs). 25-Hydroxycholesterol proteins and activation of of Bcl-2 expression down-regulation through caspases.

Down-regulates cholesterol biosynthesis in cultured Smith-Lemi- in cultured biosynthesis cholesterol Down-regulates

store at: store sealed ampule

element binding regulatory the cleavage of sterol Suppresses

Sealed ampule Do not confuse with dihydrocholesterol.

Lyco

25-Hydroxy 25-Hydroxy

7-Dehydro 7-Dehydro    Cholesterol Biosynthesis 8 sigma.com   Pravastatin sodium store at: store at: HMG-CoA reductase inhibitor Mevastatin

Inhibits myoblastformation. Cholesterol loweringdrug Competitive, water-soluble 3-hydroxy-3-methylglutaryl coenzyme A widelyusedmethodtoinduceadecrease incholesterol invivo A (HMG-CoA)reductase inhibitor. Inhibitscholesterol synthesis δ simvastatin beingthemostpotentoftheseproducts. are listedinincreasing order ofLDLloweringpotencywith to specificallyinhibitHMG-CoAreductase. Thefollowingstatins Statins are aclassofhypolipidemicagentsthathavebeenshown biosynthesis anditsdownstream effects istointroduce statins. heptanoic acid sodium

Eptastatin sodium Compactin C C solubility solubility

DMSO...... 20mg/mL

Statins ,6-tri 23 23 P4498-25MG M2537-5MG ≥ ≥

H H 98% (HPLC) 95% (HPLC) 35 34 hydroxy-2-methyl-8[(2S)-2-methyl-1-oxo

O O (K

7 5 E E

H ethanol...... 25mg/mL i ~1nM).

; FW 390.51

ML-236B 2

O...... 19 mg/mL FW 446.51

;

( β R

, δ [73573-88-3] R

,1 [81131-70-6] S

,2 S

,6 S

,8 S

,8a

)-1,2,6,7,8,8a-Hexa butoxy l]-1-naph thalene- hydro- 25 mg β 5 mg ,  

Simvastatin Mevinolin from store at:

Cholesterol loweringdrugandcompetitiveinhibitorofHMG-

store at:

Simvastatin isaspecificinhibitorofHMG-CoAreductase, the remain toberesolved. and iosprenylation ofsignalingproteins intheseobservations between drugresistance, regulation ofthemevalonatepathway, myeloma cells.Theroles ofPgpandCYP3A,possibleconnection anticancer agentssuchasdoxorubicinandinducesapoptosisin substrate ofPgpandCYP3A.Itincreases cellularresistance to the production ofcholesterol andisoprenoids. Thisproduct isa Blocks theproduction ofmevalonate,acriticalcompoundin CoA reductase, aratelimitingenzymeincholesterol synthesis. numerous otherdiseasestates. lowering cholesterol levelsandtoinvestigateitspotentialrole in in order toelucidateothercellulareffects ithasinadditionto with otherstatins,isbeinginvestigatedatthemolecularlevel for prevention and treatment ofatherosclerosis, simvastatin,along lovastatin (CatalogNumberM2147).Besidesbeingusedclinically to usewithNaOHinEtOHtreatment. Itisasyntheticanalogof to thecorresponding ß-hydroxyacid, andcanbeactivatedprior levels. Simvastatinisalactonethatreadily hydrolyzed lipoproteins andtriglycerides,raiseshigh-densitylipoprotein of hypercholesterolemia, asitreduces levelsoflow-density an earlystepincholesterol biosynthesis.Itisusedinthetreatment enzyme thatcatalyzestheconversionofHMG-CoAtomevalonate,

thalenyl esterbutanoicacid dimethyl-8-[2-(tetra C Lovastatin

solubility C 25 24 M2147-25MG S6196-25MG S6196-5MG ≥ ≥

H H 98% (HPLC) 98% (HPLC) 38 36

O O 5 5 E E

DMSO......

; FW 418.57 FW 404.54

6- α

-Methyl hydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1-naph- Aspergillus

compactin [79902-63-9] [75330-75-5]

;

Mevacor

; sp.

2-Methyl-1,2,3,7,8,8a-hexa

; ≥

Monacolin K

20 mg/mL hydro-3,7- in vivo 25 mg 25 mg 5 mg

Blocking Absorption of Dietary Cholesterol 9

sigma.com 1 g 5 g 1 g 5 g 10 g 25 g 1 mg 5 mg 10 mg 25 mg ;

-ol -ol β β ;

-methyl- β -ol β -ol tan-3 β es Stigmastan-3

;

ten-3 -ol). -chol es β α 5,22-Stigmastadien-3

;

-sitosterol -sitosterol

β -Ethyl-5

diene α ta 24

-Methyl-5-chol -Methyl-5-chol Dihydro-

α -ol; ]-ergost-5-en-3 ;

β S 24

[19466-47-8] [474-62-4] [83-48-7] ;

(soybean), ~65% (soybean),

-ol tan-3 β es -ol; 24[ -Sitostanol β

β -chol ol ;

α FW 416.72 FW 400.68 FW 412.69 Glycine max

B ester ester O O O

52 48 48 )-Ergost-5-en-3 -Ethyl-5 H H H

R -Hydroxy-24-ethyl-5,22-chol -Hydroxy-24-ethyl-5,22-chol α 95%

~95% from from S2424-5G S2424-10G S4297-1G C5157-1MG C5157-5MG C5157-10MG C5157-25MG S2424-1G S2424-25G S4297-5G

C-NMR to contain ~35% dihydrobrassicasterol (24 dihydrobrassicasterol C-NMR to contain ~35% -Sistostanol 29 28 29 β Stigmasterin C

24 24( 3 C C

β 13

5-cholesten-3

store at: store

Plant sterol. May lower absorption of dietary cholesterol. of dietary cholesterol. May lower absorption Plant sterol. and GC, but has been shown by Appears ~98% by HPLC Stigmastanol Camp Stigmasterol  

 1 mg 5 mg ;

10 mg 25 mg 10 mg 100 mg ol ter ter es chol -Ethyl β 24

;

stigmasterol stigmasterol

[83-46-5] 97%

22,23-Dihydro 22,23-Dihydro

≥ ;

95% -ol ≥ β

sterol sterol FW 414.71 fuco

B B O

50 H

from soybean, from synthetic, S9889-1MG S1270-25MG S1270-100MG S9889-5MG S9889-10MG S1270-10MG -Dihydro -Dihydro -Sitosterol -Sitosterol 29 Blocking Absorption of Dietary Cholesterol Dietary of Absorption Blocking 5-Stigmasten-3 C

β α

sources that are rich in fat content. A healthy adult only needs to A healthy adult only rich in fat content. that are sources Obtaining requirement. 30% of the daily cholesterol ingest about can lead to cholesterol dietary from than this amount more and serious health risks. levels cholesterol increased the small is absorbed within the lumen of Dietary cholesterol in the liver interact cholesterol from intestine. Bile salts produced a biliary micelle that is transported with phospholipids to produce in the lumen is Dietary cholesterol via bile into the lumen. these micelles and together with the easily incorporated into can now be absorbed into the biliary cholesterol present already of the lumen. The micelles that make up the walls enterocytes know as Niemann-Pick C1 Like 1 enter the cell by a channel can either the cholesterol (NPC1L1). Once in the cells protein the lumen or it can be esterified for be pumped back out into transport within chylomicrons. has become cholesterol the absorption of this dietary Preventing and Plant sterols research. related in cholesterol a key area inhibitors of cholesterol stanols have been shown to be effective and plant sterols absorption. Ingested as part of a normal diet, They actually to cholesterol. very similar in structure stanols are to the biliary than cholesterol binding affinity have a stronger and the sterols micelles that aid in absorption. Because of this the micelles thus preventing from stanols can displace cholesterol its absorption. the absorption of the biliary micelles inhibitors that block Recently, have also been used to block the uptake of into the enterocytes dietary cholesterol. store at: store store at: store

Plant derived estrogen Plant derived estrogen

Synthetic form of a plant derived estrogen Synthetic form of a plant derived estrogen

 

Dietary cholesterol is obtained from foods derived from animal animal foods derived from from is obtained Dietary cholesterol 10 Cholesterol Esterification sigma.com To more efficiently transportbothdietaryandsynthesized Figure 1. stream. allowing formore efficient cholesterol transportthrough theblood lipoproteins. Thisvastlyincreases thecapacityoflipoproteins, esters more cholesterol canbepackagedintotheinteriorof surface oftheparticle.Byconvertingcholesterol tocholesteryl can betakenupbylipoproteins, butisconfinedtotheouter cholesterol, itisconvertedtocholesterylesters.Free cholesterol Cholesterol Esteriﬁ Figure 2.

Lecithin:Cholesterol acyltransferase(LCAT) isfoundinperipheraltissuesandutilizes phosphatidylcholine asthesource ofa Acyl-coenzyme A:cholesterol acyltransferase2(ACAT2) isfoundintheliverand intestine,andutilizesacyl-CoAasthesource R OO OO R H R O 3 C O O CH CH N O 2 O O POH 3 CH O CH 2 R 3 H N cation O C CH 2 CH CH 2 3 H N LCAT H O C H H O OH CH 3 A C CAT O CH H CH C CH R 3 CH 3 3 2 lipids inplasma. Inhibiting theseenzymesisonewayofloweringthecirculating chains. LCAT usesphosphatidylcholinewhileACAT usesacyl-CoA. LCAT andACAT alsodiffer inthesources theyusefortheacyl peripheral tissues(see predominantly bylecithin:cholesterol acyltransferase(LCAT) inthe conversion ofcholesterol tocholesterylesteriscatalyzed conversion dependingonthelocationofreaction. The Distinct enzymescatalyzethecholesterol tocholesterylester intestine andliver(see cholesterol acyltransferase2(ACAT2), whichisfoundinboththe absorbed byenterocytes isesterifiedbyacyl-coenzymeA: O O OH P CH O 3 H O OH P OH H H O HO 3 CCH CH Figure 1 Figure 2 CH H CH H 3 O O H POH 2 3 ). Inthelumen,dietarycholesterol ). ACAT1 isfoundinalltissues. OH H N H N N NH cyl chains. 2 N of acylchains. OH 3

Cholesterol Esterification 11

sigma.com 1 g 1 g 1 g 25 g 10 g yl 100 mg 250 mg 100 mg 250 mg 500 mg ter es -Hydroxy- β Chol

;

oate dienoate decen deca tene 3-linoleate tene 3-linoleate es -9-octa yl ester yl octa cis

ter yl ter es

es ter es Chol

;

Chol

noate -Hydroxy-5-chol -Hydroxy-5-chol ;

β [601-34-3] [35602-69-8] [604-33-1] [303-43-5] 3

deca ;

Oleic acid chol

;

-yl octa -ol 3-linoleate -ol 3-linoleate -ol 3-oleate

dienoate β β β yl palmitate yl stearate yl linoleate yl linoleate yl oleate FW 625.06 FW 653.12 FW 649.08 FW 651.10 deca

B B B tene 3-palmitate ten-3 ten-3 ten-3 tene 3-oleate ter ter ter ter 2 2 2 2 es es es es O O O O

es es es es es 76 80 76 78 98% (HPLC; detection at 205 nm) at 205 98% (HPLC; detection nm) 98% (HPLC; detection at 205 98% (HPLC; detection at 205 nm) 98% (HPLC; detection at 205 H H H H

≥ ≥ ≥ 96% C6072-1G C6072-10G C79409-25G C0289-100MG C0289-250MG C0289-1G C9253-100MG C9253-250MG C9253-500MG C9253-1G 43 45 45 45 9,12-octa C 5-Chol C 5-Chol 5-Chol C 5-Chol C 5-chol

(LDL).

ester in low density lipoprotein The most abundant cholesteryl store at: store store at: store at: store

Sealed ampule Chol Chol Chol Chol     es- 25 g 50 g azine 5 mg 5 mg 100 g ;

25 mg 10 mg 25 mg 100 mg 500 mg

;

amide -Hydroxy-5-chol- phenyl)piper enoate -Hydroxy-5-chol -Hydroxy-5-chol β 3

β ;

tetra ;

ethyl]propan ethyl]propan ate

= 63-88 nM. 50 yl eicosa -(3,4-dimethyl ′

yl capro yl capro -ol 3-acetate ter β es ter

phenyl)-1-phen phenyl)-1-phen Chol

ten-3 ;

es Chol

;

[78934-83-5] oxycinnamoyl)-N [214265-97-1] 5-Chol

noate

;

[1062-96-0] [604-34-2] [604-35-3] -[2-(4-methyl -[2-(4-methyl B

-octyl

N noate tene 3-arachidonate tene n es es silyl]- -ol 3-hexa -ol 3-arachidonate

tore at: tore

FW 465.79 β β 95.0% (HPLC) FW 508.69 O...... insoluble

≥ noate yl hexa yl arachidonate yl acetate

FW 484.80 FW 673.11 FW 428.69 4 B DMSO...... >8 mg/mL H

E ten-3 ten-3 ter ter ter 2 2 2 2 dimethyl es es O O O N NOSi

es es es 56 76 48 44 47 95% (HPLC; detection at 205 nm) 95% (HPLC; detection at 205 H H H H H

-Hydroxy-5-chol -Hydroxy-5-chol -Acetoxy-5-chol ≥

purum, >98% (HPLC) >98% (HPLC) tene 3-acetate C6524-25G C8753-500MG C6524-100G C8753-10MG C8753-25MG C8753-100MG 26750-50G-F Y0628-5MG S9318-5MG S9318-25MG 33 47 29 31 30 β β DMSO...... 16 mg/mL DMSO...... 16 SA 58-035

solubility solubility C C 5-Chol 5-Chol 3 C N-(3,5-Dimethoxy-4-

3-[Decyl tene 3-hexa C

C 3

ship: dry ice s

Acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor. inhibitor. acyltransferase (ACAT) Acyl-CoA:cholesterol Enzyme target for lipid-lowering drugs IC

Acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor. inhibitor. (ACAT) acyltransferase Acyl-CoA:cholesterol

store at: room temp at: room store

Sandoz 58-035 Sandoz 58-035

store at: store Chol

Chol

Chol YIC-C8-434

store at: store     12 Cholesterol Esterification sigma.com  

Chol One unitwillhydrolyze 1.0µmoleofcholesteryloleateto Partially purified

One unitwillhydrolyze 1.0µmoleofcholesteryloleateto Contains potassiumphosphatebuffer saltsandstabilizer. Chol store at: store at: presence oftaurocholate. cholesterol andoleicacidperminat pH7.0at37°Cinthe protein 30-65% composition

presence oftaurocholate. cholesterol andoleicacidperminatpH7.037°Cinthe Protein ~70% composition Protein determinedbybiuret. Cholesterol istransportedthroughout thebodyinformof phosphatidylcholine, phosphatidylserine enolase, Hg Inhibitors: BisphenolAmethacrylate,diisopropylfluorophosphate, phosphatidylethanolamine, sodiumtaurocholic acid kinase typeII,ethanol,methanol, Activators: BisphenolAdiglycidylether, cAMP-dependentprotein Optimum pHrange:6-8 esters ofcholesterol. Enzyme responsible forthehydrolysis ofmanythefattyacid [9026-00-0] free cholesterol intotheenterocyte. cholesterol intomixedmicellesandaidsinthetransportof Cholesterol esterasealsocontributestotheincorporationof short chainsaturatedfattyacids. chain andunsaturatedfattyacidestersatagreater ratethan cholesterol andfattyacids.Cholesterol esterasehydrolyzes long- hydrolysis ofthesestored cholesterol estersyieldingbioavailable as cholesterol esters.Theenzymecholesterol esterasecontrols the cholesterol esters.Excesscholesterol isalsostored intracellulary Chol Sterol-ester acyl

Cholesterol Esterase C9464-100UN C9464-25UN C3766-500UN C3766-100UN activity: activity:

es es es ter ter ter B B ol Ester ol Ester ol Ester ≥ ≥ 2+

200 units/g protein 200 units/g 15,000 units/g protein 15,000 units/g

, sodiumfluoride,phosphatidicacid,

hydrol ase from hog pancreas hog ase from ase from porcine pancreasporcine ase from ase from bovine pancreas bovine ase from ase n -butanol, phosphatidylcholine,

500 units 100 units 100 units 25 units Diethyl    Chol

store at: Contains potassiumphosphateandTRITON

store at: store at:

One unitwillhydrolyze 1.0µmoleofcholesteryloleateto Selective, potentcholesterol esteraseinhibitor. Blocks

Protein ~20% composition presence oftaurocholate. cholesterol andoleicacidperminatpH7.037°Cinthe cholesterol. IC esterase are anticipatedtolimittheabsorptionofdietary the mitochondriaofsteroidogenic cells.Inhibitorsofcholesterol steroidogenesis primarily bypreventing cholesterol transportinto C DEUP solubility 14 C1403-100UN C1403-25UN C9281-500UN C9281-100UN D7692-25MG D7692-5MG activity: activity: >98% (HPLC)

H 17

; es

UBP 6 ter B B B umbelli

DMSO...... 5mg/mL

ol Ester ≥ ≥ FW 312.25 200,000 units/g protein 200,000 units/g protein 10,000 units/g

50 =11.6µM.

feryl phos ase from ase from

[897-83-6] phate Pseudomonas ﬂ

® X-100.

uorescens

100 units 500 units 100 units 25 units 25 mg 5 mg Cholesterol Transport 13

sigma.com

). Figure 3 Figure Transmembrane Region O-Linked Glycans Ligand Binding DomainLigand Binding N-Linked Glycans Cytoplasmic Domain . LDL Receptor. Figure 3 Figure ATP-dependent proton pumps lower the pH inside the vesicle proton ATP-dependent After loss of the its receptor. in dissociation of LDL from resulting in peptide resulting clathrin coat, the vesicles fuse with lysozomes can The LDL receptor and cholesteryl ester enzymatic hydrolysis. and to the cell membrane. Insulin, triiodothyronine, be recycled of LDL the regulation dexamethasone have been shown to affect mediated endocytosis. receptor the smallest of the lipoproteins (HDL) are High density lipoproteins and most dense. HDL contains several types of apolipoproteins apo-CI, II & III, apoD and apoE. HDL including apo-AI, II & IV, phospholipids, cholesteryl esters, and contains mostly protein, in the liver rich particle as a protein HDL is produced cholesterol. of Apo-CI & II and source and intestine, and serves as a circulating particle accumulates cholesteryl The HDL protein ApoE proteins. by lecithin:cholesterol esters by the esterification of cholesterol apo-AI on HDL. HDL is activated by LCAT acyl-transferase (LCAT). cell membranes and can transfer from cholesterol can acquire cholesteryl esters to VLDL and LDL via the transferase activity of by is removed cholesterol apoD. HDL can return liver where to the transport, thus, serving as a scavenger of free cholesterol reverse cholesterol. facilitates the The cytoplasmic domain of the LDL receptor of the membrane. regions formation of coated pits; receptor-rich apo-B100 recognizes The ligand binding domain of the receptor formation of a clathrin-coated vesicle that in the on LDL, resulting the inner surface of the cell membrane (see buds from

HDL LDL IDL Cholesteryl Esters Cholesterol Triglycerides Phospholipids Apo-B100 Chylomicron VLDL ). VLDLs contain triacylglycerols, ). VLDLs contain triacylglycerols, Figure 2 Figure ). Chylomicrons predominately transport triacylglycerols transport triacylglycerols predominately ). Chylomicrons Low Density Lipoprotein. Chylomicron. Figure 2. Figure some cholesterol and cholesteryl esters and the apoproteins; apo- and cholesteryl esters and the apoproteins; some cholesterol exist to remove B100, apo-CI, apo-CII, apo-CIII, and apoE. VLDLs the liver and distribute and cholesteryl esters from triacylglycerols the circulating As VLDLs move into the body. them throughout converted first to intermediate density lipoproteins plasma they are (LDL). Lipoprotein (IDL) and then into low density lipoproteins both the of fatty acids from the majority lipase serves to remove while density of the lipoproteins the VLDL and IDL, thus increasing and cholesteryl ester concentrations. maintaining cholesterol all apolipoproteins of fatty acids and the loss of The removal the primary in LDL. LDLs are except apoB-100 and apo(a) results LDL can for delivery to all tissues. plasma carriers of cholesterol mediated be absorbed by the liver and other tissues via receptor endocytosis. Figure 1. Figure dense smaller and more (VLDL) are low density lipoproteins Very (see than chylomicrons Cholesterol Transport Cholesterol

packaged for transport within the plasma. The particles that plasma. The particles that transport within the packaged for for esters, and triglycerides cholesteryl package cholesterol, five main classifications are There lipoproteins. called transport, are the The higher density. based on their size and of lipoproteins to lipid content the higher the density. ratio of protein 1 Figure as fatty acids, but also deliver dietary to adipose tissue and muscle Once in the lumen to the liver. taken up by enterocytes cholesterol to the adipose have been delivered most of the triacylglycerols including of the lipoprotein, remnants tissue and muscle, the to, and taken then delivered apoE, and apo-B48 are cholesterol, remnant interaction with the chylomicron the liver through up by, receptor.

The largest and least dense lipoproteins are chylomicrons (see chylomicrons are lipoproteins The largest and least dense

Since cholesterol is a water-insoluble molecule it must be molecule it must is a water-insoluble Since cholesterol 14 Cholesterol Transport sigma.com  

Apo Solution in10mMammoniumbicarbonate. Solution in10mMammoniumbicarbonate Apo

mol wt17.38 kDa Major protein componentsinhighdensitylipoprotein (HDL) Apo-AI comprises~70%oftheprotein moietyinHDL.Itisa Forms dimersinaqueoussolutionswithanassociationconstantof Apo A-IIconstitutes~20%oftheprotein moietyofhighdensity Unlike ApoA-I,A-IIpossessesnoactivatingproperties for of ~35%in may beinverselyrelated totheriskofcoronary disease. Apo-AI levelsinnormalplasmaare 90-130 mg/dl.Apo-AIlevels esterification inplasma. acyltransferase (LCAT), whichisresponsible forcholesterol conformation change.Apo-AIactivateslecithin:cholesterol In aqueoussolution,Apo-AIshowsself-associationwithminor α content of and twominorisoforms(pI5.535.46).Apo-AIshowsahigh 28.3 kDa.Theprotein ismadeupofonemajorisoform(pI5.6) the N-terminalresidue. Themolecularmassisreported tobe glutamic acidastheC-terminalresidue andasparticacidas single polypeptidechainconsistingof245aminoacidswith ship: dryicest 5 ×10 lipoprotein (HDL). found thatApoA-IIinhibitstheactivationofLCAT byApoA-I. lecithin:cholesterol acyltransferase(LCAT). Furthermore, itwas ship: dryices Apo A-I Apo A-II Apo-AI (28kDa) Apo(a) (300-800kDa) Apo-H (50kDa) Apo-E (34kDa) Apo-D (20kDa) Apo-CIII (8.8kDa) Apo-CII (8.8kDa) Apo-CI (6.6kDa) Apo-B100 (513kDa) Apo-B48 (241kDa) Apo-AIV (46kDa) Apo-AII (17.4kDa) Apolipoprotein CompositionofLipoproteins -helix structure seemtoberesponsible forlipidbindingcapacity. A0722-1MG A0722-.5MG A0972-1MG A0972-.5MG >95% (SDS-PAGE) ≥ 85% (SDS-PAGE) lipo lipo 4 M

protein A-I from humanplasma protein A-II from humanplasma -1 α . Theassociationisaccompaniedbyanenhancement

α -helix structure. Theamphipathicregions inthe tore at: ore at: -helical content. B B

hl-irn LL D D HDL LDL IDL VLDL Chylo-micron 0.5 mg 0.5 mg 1 mg 1 mg X X X XX XX XX XX X X    Apo Apo Apo Delipidated withsodiumdeoxycholate. Lyophilized from buffer containing10mMsodiumdeoxycholate, Lyophilized from 10mMNH store at: store at: store at: 0.05 Msodiumcarbonate,andchloride,pH10.0 average molwt8.8 kDa Activates lipoprotein lipase average molwt6.6 kDa Very lowdensitylipoprotein. Interferes directly withfattyaciduptakeandisthemajorplasma

inhibitor ofcholesterol estertransferprotein. A7910-50UG A7785-100UG A5353-.5MG ~95% >95% (SDS-PAGE) >95% (SDS-PAGE)

lipo lipo lipo B B B protein B from humanplasma protein C-II from humanplasma protein C-I from humanplasma XXX

X X X X 4 HCO

3 , pH7.5

X X 0.5 mg 100 µg 50 µg

Cholesterol Transport 15

sigma.com 1 vial 1 mg 5 mg 50 µg 10 mg 10 mg VLDL

;

protein protein protein protein protein protein protein, the major component of protein, -Lipo

a β

;

cells s disease, which it delivers to the microglia, to the microglia, s disease, which it delivers ′ E E protein protein Low density lipo

;

Very low density lipo Very

;

tore at: tore

store at: store protein protein protein E4 human protein protein protein E E E B -Lipo High density lipo

-lipo protein, low density from human plasma from low density protein, human plasma from very low density protein, protein, high density from human plasma from high density protein, lipo

β ;

95% (SDS-PAGE) 95% (SDS-PAGE) 95% (SDS-PAGE) 95% (SDS-PAGE) β ;

≥ ≥

>95% (SDS-PAGE) >95% (SDS-PAGE) recombinant, expressed in baculovirus infected expressed recombinant, L7527-1MG L1567-10MG L8039-10MG L8292-1VL L7914-5MG A2456-50UG Pre- LDL HDL

ship: wet ice ship: wet ice s increasing plasma lipid levels by regulates It plasma lipoproteins. triglycerides and cholesterol. of particles rich in the degradation apolipoprotein and particles containing It binds to LDL receptors E4 and also binds amyloid- plaques in Alzheimer of brain. the major scavenger cells pH 7.4 protein mg vial of ~5 pH 7.4

E family of of the apolipoprotein E4 is a member Apolipoprotein Spodoptera frugiperda for binding to the human Apo B/E (LDL) low density lipoprotein receptor. store at: store store at: store at: store store at: store Lyophilized from a solution of 0.15 M NaCl and 0.01% EDTA, a solution of 0.15 M NaCl and 0.01% EDTA, from Lyophilized vial of ~10 mg protein (modified Lowry) mg protein vial of ~10 pH 7.4 Solution in 150 mM NaCl and 0.01% EDTA, pH 7.4 Solution in 150 mM NaCl and 0.01% EDTA, mg protein vial of 10 pH 7.4 Solution in 150 mM NaCl and 0.01% EDTA, competes with iodinated human Apo E4 Human recombinant bicarbonate. Solution in 0.7 M ammonium

Lipo Lipo Apo Lipo a solution of 0.15 M NaCl and 0.01% EDTA, from Lyophilized    

50 µg 50 µg 8 100 µg -amyloid -amyloid β β protein, the major protein, β s disease, which they deliver ′

B protein, the major component of plaques protein, β s disease. ′ s disease, which it delivers to the microglia, the s disease, which it delivers to the microglia,

′

21 cells cells Sf Sf protein E3 human E3 protein protein E2 human human E2 protein protein C-III from human plasma human from C-III protein B B lipo lipo lipo 95% (SDS-PAGE), recombinant, expressed in baculovirus expressed recombinant, 95% (SDS-PAGE), 90% (SDS-PAGE), recombinant, expressed in baculovirus expressed recombinant, 90% (SDS-PAGE), ≥ ≥ A2331-50UG A2673-50UG A3106-100UG Apo E3 Apo E2 Apo C-III

levels by increasing plasma lipid It regulates plasma lipoproteins. and cholesterol. the degradation of particles rich in triglycerides and particles containing apolipoprotein It binds to LDL receptors, E2 bind amyloid- in Alzheimer E3, to apolipoprotein major scavenger cells of brain. Compared levels E2 is associated with lower plasma LDL apolipoprotein against the development of atherosclerosis. and may protect risk E2 also appears to be associated with reduced Apolipoprotein for Alzheimer

It is an inhibitor of of triglyceride-rich lipoproteins. metabolism in higher levels of Apo C-III result lipase (LPL). Reduced lipoprotein triglycerides into adipose tissue. plasma fatty acid uptake from

component of plaques in Alzheimer the major scavenger cells of brain. to the microglia, for binding to the human Apo B/E (LDL) low density lipoprotein Apo E also binds to Human recombinant receptor. peptide in a soluble binding assay.

plasma lipid levels It regulates E family. allele of the apolipoprotein the degradation of particles rich in triglycerides by increasing and is involved in the It binds to LDL receptors and cholesterol. ApoE-containing particles in development of atherosclerosis. fluid bind amyloid- plasma and cerebrospinal infected

Apolipoprotein E2 is a member of the apolipoprotein E family of the apolipoprotein E2 is a member of Apolipoprotein

mol wt ~34 kDa mol wt ~34 infected Apolipoprotein C-III (Apo C-III) is central in regulating the regulating (Apo C-III) is central in C-III Apolipoprotein

ship: dry ice store at: ship: dry ice store store at: store store at: store Human recombinant Apo E3 competes with iodinated human Human recombinant Solution in 0.7 M ammonium bicarbonate. kDa mol wt ~34 for binding to the human Apo B/E (LDL) low density lipoprotein Apo E also binds to Human recombinant receptor. mol wt 8.75 kDa mol wt 8.75 peptide in a soluble binding assay. Apolipoprotein E3 is a major plasma lipoprotein and the dominant E3 is a major plasma lipoprotein Apolipoprotein

Human recombinant Apo E2 competes with iodinated human Apo E2 competes with iodinated Human recombinant Solution in 0.7 M ammonium bicarbonate.

Apo Apo

Solution in 10 mM ammonium bicarbonate. Solution in 10 mM ammonium Apo   16 Cholesterol Transport sigma.com  Selectively increases Apolipoprotein A-Ilevels. Gemfibrozil Fenofibrate reducing cholesterylestertransferprotein expression. L A numberofcompoundshavetheabilitytoaffect levelsof

with inhibitorsofcholesterol biosynthesis. greatly reduced whenthesecompoundsare usedincombination lipoproteins circulating intheblood.Riskofatherosclerosis canbe C C 2,2-Dimethyl-5-(2,5-dimethyl 2-[4-(4-Chloro

Lipoprotein Regulation ipid regulating drug.Increases highdensitylipoprotein levelsby 20 15 G9518-25G G9518-5G F6020-100G F6020-25G F6020-5G ≥

H H 99% 22 21

O ClO 3

FW 250.33 FW 360.83 benzoyl)phen Proteinase K,AlkalinePhosphataseandLeupeptin Proteomics, andCellCulture Introducing the and technicalinformationyouneed 2000+ To reserveyour copy, New! LifeScienceWeb Tool Guide

New Sigma

[25812-30-0] [49562-28-9] oxy]-2-methyl phen Products oxy)pentanoic acid Your workisunique...innovative...groundbreaking New ®

LifeScienceCatalogistheperfectmatch propan

supportingGenomics,Functional Cell Biology, GeneExpression, BioUltraProteins visit oic acidiso

sigma.com/sigmacat1 The New2008-2009 (Available January2008) propyl ester – learn abouttheoptimalonlinetool for ﬁ –learn –risk-free, high-purityproteins, including 100 g 25 g 25 g 5 g 5 g  Enhances selectiveuptakeofHDL-associatedcholesterylesters. Probucol Nico Decreases hepaticproduction ofvery-lowdensitylipoproteins and apolipoprotein B.

C Niacin C

Vitamin B Vitamin 31 6 P9672-50G N4126-1KG N4126-500G N4126-100G N4126-5G ≥

H 98% 5 48 NO tinic acid

;

Pellagra preventive factor 2 2

S 3 2

FW 123.11

FW 516.84

[59-67-6]

[23288-49-5]

; ndingtheproducts

3-Pico

linic acid

;

Pyri dine-3-carboxy lic acid 500 g 100 g 50 g 1 kg 5 g

; Bile Acids 17

sigma.com A Co - S - C O OH 7-Hydroxycholesterol Cholesterol Propionyl-CoA H ) Bile acids are then delivered to the intestines where to the intestines where delivered then ) Bile acids are 2 CoA-SH NADPH + H Chenodeoxycholyl-CoA 2 O NADP Figure 1 Figure NADPH + H HO 2 O 7-hydroxycholesterol. 7-hydroxycholesterol is converted to one of 7-hydroxycholesterol 7-hydroxycholesterol. acid. and chenodeoxycholic bile acids, cholic acid the two primary (see lost during bile acids are of lipids. Some they aid in absorption to the acids delivered a majority of bile however, this process; by absorption in the ileum and returned to recycled intestine are bile acids are Back in the liver glyco- and tauroconjugate the liver. gall bladder for storage and use again as formed and moved to the digestion aids. 2 Multiple Reactions Multiple O α 7 -Hydroxylase CoA - 2 CoA-SH NADPH + H S - C O Propionyl-CoA HO HO OH OH OH H ). Activation of LXRs by specific ). Activation of LXRs by β Cholyl-CoA and LXR- HO α Bile Acid Biosynthesis Pathway. -hydroxylase, which is the rate limiting enzyme in the pathway -hydroxylase, α Bile Acids Bile Figure 1. Figure cholesterol unwanted two purposes: to remove Bile acids serve in the intestine. Bile aid in lipid digestion the body and to from is tightly production in hepatocytes and this synthesized acids are transcription factors, Liver nuclear by the receptor controlled X Receptors (LXR- oxysterol derivatives leads to the regulation of bile acid synthesis the regulation derivatives leads to oxysterol the activation of transport. LXRs control cholesterol and reverse 7 This enzyme converts cholesterol into for bile acid biosynthesis. The liver excretes excess cholesterol in the form of bile acids. cholesterol excess The liver excretes 18 Bile Acids sigma.com     Deoxy Bile Acid Dehydro Cheno Glyco Bile Acid Bile Acid Cholic acid Bile Acid

Bile Acid

methyl)amide C 7-Deoxy C 5 Cheno Cholanic acid Cholyl C C C solubility β 24 24 24 26 24 C1129-1KG C1129-500G C1129-100G C1129-25G D2510-500G D2510-100G D2510-10G D3750-25G C9377-25G C9377-5G C9377-100MG G2878-25G G2878-5G G2878-1G G2878-500MG G2878-100MG from oxorsheepbile, synthetic, ≥ ≥

-Cholanic acid-3,5,12-tri

H H H H H 99% (TLCandtitration) 98% 34 40 40 43 40 gly

O O O O NO diol cholic acid hydrate deoxy cholic acid cholic acid 4 5 4 5 cine

ethanol...... 10mg/mL 6

;

·xH cholic acid FW 392.57 FW 402.52 FW 392.57 FW 408.57

5 β

;

3 -Cholanic acid-3

3 N

≥ 2 cholic acid α α

O -(3 97% (TLC) ,7 ,7

; α α α

Desoxy FW 465.62(Anh) ,7 ,12 ,12

α α [83-44-3] [81-23-2] [474-25-9] [81-25-4] ,12 α -Tri -Tri cholic acid one α hydroxy-5 hydroxy-5 -Tri α ≥

;

3,7,12-Trioxo-5 hydroxy-24-oxocholan-24-yl)-gly ,7 98% α

-diol

;

3 β β [475-31-0] α -cholan-24-oic acid

; -cholanic acid ,12

3 α ,7 α -Dihydroxy-5 α β -Dihydroxy-5 -cholanic acid

β N -cholanic acid β -(carboxy- -cholanic acid cine 500 mg 100 mg 100 mg 500 g 100 g 500 g 100 g 25 g 25 g 25 g 10 g 1 kg 25 g 5 g 1 g 5 g    Sodium cholate hydrate Sodium cheno Litho

Bile Salt Bile Acid Solubilizes fatsforabsorptionintheintestines. Sodium deoxy Bile Salt

 salt acid sodium

Dihydroxy-5

5 C C Cholalic acid sodium salt Cheno 5 7-Deoxy C

5 β β β 24 24 24 24 C8261-1G C8261-500MG L6250-25G L6250-10G D6750-500G D6750-100G D6750-25G D6750-10G C1254-5KG C1254-1KG C1254-500G C1254-100G C1254-25G from oxorsheepbile, ≥ ≥ ≥

-Cholanic acid-3 -cholan-24-oic acid -Cholan-24-oic acid-3

H H H H 97% 97% (titration) 97% (titration) 39 40 39 39 cholic acid

NaO O NaO NaO deoxy cholic acid sodium salt 3

4 5 4 FW 376.57

·xH

β cholic acid sodium salt -cholanic acid sodium salt FW 414.55 FW 414.55 2

O deoxy cholate

α ,7 FW 430.55(Anh)

α [434-13-9] -diol

; α

cholate

3 -ol [2646-38-0] [302-95-4] α

;

; ,7

3 ; ≥ α α α

Desoxy 99% ,12 ,7 -Hydroxy-5

; α α

Chenodesoxy

-Dihydroxy-5 -Tri

cholic acid sodium salt

[206986-87-0] hydroxy-5

β -cholanic acid β cholic acid β -cholanic acid -cholan-24-oic

;

Cheno α

3 -Hydroxy- α ,12 diol 500 mg α 500 g 100 g 500 g 100 g - 25 g 25 g 10 g 25 g 10 g 5 kg 1 kg

; 1 g Bile Acids 19

sigma.com ;

1 g 5 g 1 g 5 g 1 g 5 g 25 g 25 g 50 g 100 g 250 mg 500 mg 100 mg 500 mg 0.05% 0.005% 0.005% 0.001% ≤ ;

0.0005% ≤ ≤ ≤ ≤ sulfonic acid

methyl)

-cholan-24-oic acid

β )......

2- 4 -(carboxy N hydroxy-5 [863-57-0] [345909-26-4]

/taurocholate cotransporter /taurocholate -Tri -Tri

(+) α 0.1% hydroxy-24-oxo-5b-cholan- 0.002% [207737-97-1] ≤ hydroxy-24-oxocholan-24-yl) ,12 ≤

α cholic acid sodium salt hydrate -Tri -Tri -cholan-24-yl]amino)ethane ,7 α β α 3

0.01% Mg...... 0.05% sulfate (SO ,12 ;

0.05% Pb...... Glyco

≤ ≤ ;

α ≤ 0.0005% Fe...... 0.0005% Ca...... -cholan-24-oic acid -cholan-24-oic 0.0005% ,7 ≤ ≤ FW 487.60 (Anh) cholate hydrate β ≤ FW 537.68 (Anh) α 2-[(3a,7a,12a-Tri 2-[(3a,7a,12a-Tri

;

-(3

N 97% (TLC) sulfonic acid 2 O

FW 521.69 (Anh) ;

2 ≥ cholic acid hydrate hydrate cholic acid deoxy cholate hydrate cholate hydrate

hydroxy-5 S · xH

O...... 0.5 M at 20 °C, clear, colorless O...... 0.5 M to at 20 °C, clear,

)......

- 7 2 -Tri -Tri cine sodium salt -Dihydroxy-24-oxo-5 cholic acid sodium salt hydrate sodium salt cholic acid Na · xH SNa

ethyl)amide α 6 6 α H

gly ,12 NNaO NO NO

,12 ......

deoxy α 44 42 44

+ α 95% (TLC) 97% (TLC) 4 H H H

,7 cine sodium salt ≥ ≥ SigmaUltra, T0557-5G T0557-500MG T0557-1G T4009-250MG T4009-1G T4009-5G T4009-25G G7132-100MG G7132-500MG G7132-1G T4009-100G G7132-5G G7132-25G G7132-50G -(2-sulfo 26 -Cholyl 26 26 ile salt, physiological transport substrate for the bile salt export α Al...... chloride (Cl K...... Zn...... Cu...... NH Insoluble matter...... Phosphorus (P)...... 3 Tauro Tauro

C solubility

Sodium taurocholate hydrate Sodium taurocholate 2-([3 C

faintly yellow amide sodium amide sodium salt N 24-yl)amino]ethane

B pump/sister of Pgp (BSEP/spgp), Na (NTCP) and MRP3 into the hepatic carnalicular. N gly

Bile Salt cholic acid Synthesized from Bile Salt Bile Salt Sodium tauro Sodium tauro Sodium glyco Sodium glyco Sodium tauro    5 g 1 g 5 g 25 g 250 mg 0.05% 0.02% ;

0.05% 0.005% 0.001% 0.001% ≤ ≤ 0.001% 0.005% 0.002% ≤ 0.0005% 0.0005% 0.0005% 0.0005% ≤ ≤ ≤ 0.0005% 0.0005% ≤ ≤ ≤ ≤ ≤ ≤ ≤ cholic ≤ ≤ deoxy )...... )......

methyl)amide

2- 2- 4 4

Glyco

;

cine

-(carboxy N sulfate (SO 0.1% 0.001% ≤ ≤ 0.10 0.08 0.5% [145224-92-6] ≤ ≤ 1% sulfate (SO ≤

≤ cholic acid 0.05% Pb...... [16409-34-0] 0.002% Ba...... 0.005% Mg...... 0.002% Zn...... 0.001% K...... ≤ 0.0005% Ca...... 0.0005% Co...... 0.0005% Fe...... 0.0005% Mg...... 0.0005% Mo...... 0.0005% Cu...... 0.0005% Ca......

≤ ≤ ≤ 0.0005% Pb...... ≤ 0.0005% ≤ O at 20 °C O at 20 ≤ ≤ ≤ ≤ ≤ ≤ cholate 2 ≤ ≤ desoxy -cholan-24-oic acid FW 432.57 β 97% (TLC) 99% (titration) 99% (titration)

cholate monohydrate cholate monohydrate ≥ ≥ Glyco

deoxy O

;

FW 471.61 2

O...... 0.1 M at 20 °C, clear, colorless O...... 0.1 M at 20 °C, clear, O...... 0.1 M at 20 °C, clear, colorless M O...... 0.1 20 °C, at clear,

)......

- 5 2 2 · H

4 -Dihydroxy-24-oxocholan-24-yl)gly -Dihydroxy-24-oxocholan-24-yl)gly O...... O......

2 2 H H

α -Dihydroxy-5 H

H NNaO NaO

, , α ,12 ......

42 39

+ α 4 H H

,12 260 260

SigmaUltra, SigmaUltra, SigmaUltra, 0.1M 0.1M G9910-250MG G9910-1G G9910-5G D5670-25G D5670-5G -(3 26 24 α Al...... Al...... Cu...... NH Zn...... chloride (Cl Cd...... Sr...... Sr...... pH....7.5-9.5, 0.1 M H pH....7.5-9.5, 0.1 Cr...... Fe...... Li...... M...... K...... Bi...... Ni...... Cholic acid...... Insoluble matter...... Phosphorus (P)...... Insoluble matter...... passes filter test Insoluble matter......

solubility solubility

3 C

A A

acid sodium salt Induces apoptosis in hepatocytes; may induce DNA cleavage. Induces apoptosis in hepatocytes; may induce

Bile Salt

Sodium deoxy Sodium deoxy  Sodium glyco  20 Bile Acids sigma.com Cholic acid(C1129)25g    Chenodeoxycholic acid(C9377)100mg Lithocholic acid(L6250)1g

T0901317 ( Mixture ofsodiumcholateanddeoxycholate Bile salts Bile AcidsKit store at: Deoxycholic acid(D2510)10g Dehydrocholic acid(D3750)25g Components Bile acidsinquantitiesindicated

store at:

mol wt481.3 LXR agonistwhosetreatment results inanLXR-dependentup- A naturalproduct thatlowerscholesterol duetoitsfunctionas regulation ofABC1 geneexpression Xreceptorregulation. (FXR)modulator. Selective farnesoid in cholesterol metabolism.Playsarole incholesterol level receptor thatregulates thetranscriptionofseveralgenesinvolved an antagonistligandforbileacidreceptor, anuclearhormone

C 4,17(20)- dione solubility C

E-form...... ~5% Z 21 17 BA-1KT T2320-5MG G5168-25MG G5168-5MG B8756-500G B8756-100G B8756-50G B8756-10G >98% ~94% (HPLC) ≥ )-Guggulsterone

H H 95% (TLC) 12 28

O NSO 2 B E

cis

DMSO...... 5mg/mL 3 FW 312.45

-Pregna 9

FW 481.33 diene-3,16-dione

[39025-23-5]

[293754-55-9]

;

(17

Z )-Pregna-4,17(20)-diene-3,16-

25 mg 500 g 100 g 5 mg 5 mg 50 g 10 g 1 kit  BSEP human

Supplied asisolated The quantityoftransportedmoleculescanbedeterminedby The bilesaltexportpump(BSEP/ABCB11)belongstothefamilyof suspended in50mMHEPES-Tris, 100mM KNO Distributed forSOLVO Biotechnology, Inc. The vesiculartransportassaydeterminestheinteractionof

also bylabelingwithfluorescent orradioactive( methods suchasHPLC,LC/MS/MSseparationanddetection, important causeofcholestaticliverdisease. of geneexpression, disturbedsignaling,orstericinhibition,isan transporters suchasBSEP, duetogeneticmutation,suppression for drug-inducedcholestasis.Dysfunctionofindividualbilesalt or drugmetabolites.Thisispotentiallyasignificantmechanism canalicular membraneandisinhibitedbyanumberofdrugs energy source. BSEPisthemajorbilesalttransporterinliver transport substratesacross thecellmembraneusingATP asan the sisterofP-glycoprotein (sisterPgp).MostABCtransporters ATP-binding-cassette (ABC) transportersandhasalsobeencalled the vesicles. molecules from thesurrounding buffer andtransportstheminto out vesicles,transportofsubstratesacross themembranetakes orientation (5-10%oftotallipid).Inthecasetheseinside- contain someclosedmembranevesiclesthathaveaninside-out transporters. Membranepreparations from infectedcellsalways BSEP intomembranevesiclespurifiedfrom sucrose, pH7.4. by changesintheinitialrateof compounds withtheBSEPtransporter. Theinteractionisdetected n shi increasing therateofTCtransport compared tothe control level. vesicles. Somecompoundscanbeco-transportedwithTC, transporter, itwillblockthetransportofTCintomembrane the rateofTCtransport.Ifacompoundisaninhibitor of thetransporter, itmightcompetewithTC,thus,reducing other compoundsadded.Ifasubstanceistransportedsubstrate modulate theinitialrateofTCtransportmeasured withoutany very efficiently. Compoundsthatinteractwiththetransporter acid) tags.BSEPmediatesthetransportoftaurocholic acid(TC) ABCB11 B2436-500UL for Vesicular Transport, recombinant, expressed in

p: dryicestore at:

;

Bile saltexportpump A Sf 9 cellmembranescontaininghumanBSEP

3 H-taurocholic acidtransportby

Sf 9 cellsexpressing the

3 3 , and50mM

H-taurocholic

Sf 9 cells

500 µL The Online Resource for Nutrition Research Products Only from Sigma-Aldrich

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