DOCSLIB.ORG

Between Amphiphilic and Hydrophilic Proteins in Detergent

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Between Amphiphilic and Hydrophilic Proteins in Detergent Proc. Natl. Acad. Sci. USA Vol. 74, No. 2, pp. 529-532, February 1977 Biochemistry Charge shift electrophoresis: Simple method for distinguishing between amphiphilic and hydrophilic proteins in detergent solution (membrane proteins/protein analysis/Triton X-100/cytochrome bs/Semliki Forest virus) ARi HELENIUS AND KAI SIMONS European Molecular Biology Laboratory, Postfach 10.2209, 6900 Heidelberg, Federal Republic of Germany Communicated by John Kendrew, November 18, 1976 ABSTRACT Seventeen hydrophilic proteins and five am- MATERIALS AND METHODS phiphilic membrane proteins were subjected to agarose gel electrophoresis in the presence of a nonionic detergent (Triton The amphiphilic membrane proteins used were microsomal X-100), a mixture of a nonionic and an anionic detergent (Triton cytochrome b5 (a generous gift from Dr. C. Lussan, Talence, X-100 and sodium deoxycholate), and a mixture of a nonionic and a cationic detergent (Triton X-100 and cetyltrimethylam- France), intestinal brush border aminopeptidase (a generous monium bromide). The electrophoretic mobility of the hydro- gift from Dr. S. Maroux, Marseille, France), membrane peni- philic proteins was unaffected in the three detergent mixtures. cillinase from Bacillus licheniformis (8), and the glycoproteins However, the mobility of the hi hilic proteins shifted an- El and E2 from Semliki Forest virus membrane (9). The fol- odally in the Triton X-100-deo a e system and cathodally lowing soluble proteins were used: the trypsin-cleaved form of in the Triton X-100-cetyltrimethylammonium bromide system the brush border aminopeptidase (a gift from Drs. S. Maroux when compared to the mobility in Triton X-100 alone. The de- tergent-induced shift in mobility provides a simple, rapid, and and D. Louvard, Marseille, France, ref. 10); the polar heme- sensitive method for distinguishing between hydrophilic and containing domain of cytochrome b5 (obtained by trypsin amphiphilic proteins. cleavage of the membrane form, ref. 11); the exopenicillinase (isolated from Bacillus licheniformis, ref. 12); transferrin and In the characterization of membrane proteins it is important gamma globulin (Kabi, Sweden); lysozyme, ferritin, cyto- to be able to decide whether the proteins studied possess hy- chrome c, and myoglobin (Serva, Federal Republic of Ger- drophobic domains that anchor them to the hydrocarbon in- many); chymotrypsinogen and pancreatic ribonuclease terior of the bilayer or whether they are externally bound to the (Worthington, USA); bovine serum albumin (Calbiochem, membrane. The electrophoretic screening method introduced USA); catalase and glutamate dehydrogenase (Miles, Great in this paper is based on the fundamental difference in the in- Britain); and thyroglobulin, insulin, and ovalbumin (Sigma, teraction of hydrophilic and amphiphilic proteins with "mild" USA). Triton X-100 (Rohm & Haas, USA), sodium deoxycholate detergents such as Triton X-100 (p-t-octylphenylpolyoxy- (Schwarz/Mann, USA), and cetyltrimethylammonium bromide ethyleneg-1o). It has been shown in numerous studies that (Serva, Federal Republic of Germany) were used without pu- whereas ordinary soluble proteins and peripheral membrane rification. proteins bind little or no Triton X-100, amphiphilic membrane The agarose electrophoresis experiments were performed proteins bind large amounts (usually 80-100 mol of Triton per at room temperature (250) in 1% agarose (BioRad, USA) on glass mol of protein) when solubilized from membranes (refs. 1-5; plates (11 X 20.5 cm) as described by Weeke (13), using a for reviews see refs. 6 and 7). The bound detergent forms mi- water-cooled chamber (Behringwerke, Federal Republic of celle-like clusters around the hydrophobic domains of these Germany), paper wicks, and 1 X 11 mm sample slits. The gel proteins, and usually the proteins retain their native confor- buffer was 0.05 M glycine-NaOH, 0.1 M NaCl at pH 9.0 con- mation. Several methods have been described to determine taining 5 g of Triton X-100 or 5 g of Triton X-100 and 2.5 g of Triton X-100 binding (1-3, 5). Each of these methods can as sodium deoxycholate or 5 g of Triton X-100 and 0.5 g of such be used to differentiate between amphiphilic and hy- cetyltrimethylammonium bromide per liter. The buffers in drophilic proteins, but contrary to the method described here, the electrode chambers were the same except that in the ex- they require purified protein (usually in milligram amounts) periments with Triton X-100 no detergent was added. The and radioactive detergent. plates were first electrophoresed for 15 min at 4.5 V/cm. Instead of using Triton X-100 alone we have used mixtures Samples (12 Al) were applied and electrophoresed for 2 hr at of Triton X-100 and charged detergents. When solubilized with 4.5 V/cm unless otherwise indicated. After electrophoresis the such detergent mixtures, the amphiphilic proteins form de- gels were immediately dried under an air fan (40450) and then tergent-protein complexes containing both neutral and charged autoradiographed using Kodak Royal X-Omat film, scanned detergent molecules (see ref. 6). The net charges of the com- with a Perkin-Elmer 156 double wavelength spectrophotom- plexes are thus dependent on the charge of the detergents used, eter, or stained for protein (45 min in 0.7 g of Coomassie blue, resulting in a clear-cut difference in electrophoretic mobility 450 ml of methanol, and 90 ml of acetic acid per liter; rinsed of the amphiphilic proteins when electrophoresed in cationic for 15 min in 75 ml of acetic acid and 50 ml of methanol per and anionic detergent mixtures. The electrophoretic mobility liter). Immunoelectrophoresis was performed in agarose (1 of the hydrophilic proteins, which do not interact with the g/100 ml) gels according to Scheidegger (14), except that the detergents, remains unaffected by a change in the charge of the buffer (0.06 M sodium barbital diethylbarbitate-HCI, pH 8.7) detergents used. contained Triton X-100, deoxycholate, and cetyltrimeth- 529 Downloaded by guest on October 2, 2021 530 Biochemistry: Helenius and Simons Proc. Natl. Acad. Sci. USA 74 (1977) INSULIN BSA OVALBUMIN FERRITIN THYROGLOBULIN c2cm ORIGIN - l-l-4l - i I i CATALASE GDH TRANSFERRIN MYOGLOBIN GAMMAGLOBULIN LL Mr ORIGIN -. I1 +I i- I I I { a CHYMO- CD NO DETERGENT RIBONUCLEASE TRYPSINOGEN LYSOZYME CYTOCHROME C a TX-DOC ORIGIN -10 I~ I I i I i I I II I a TX cz3 _ TX -CTAB a_ _~ S- CO FIG. 1. Combined results from agarose gel electrophoresis of soluble proteins in the absence of detergent, in Triton X-100 (TX) and sodium deoxycholate (DOC), Triton X-100 alone, and Triton X-100-cetyltrimethylammonium bromide (CTAB). The samples applied contained 35 jig of protein in 12 Ml. Protein stain: Coomassie blue. BSA, bovine serum albumin; GDH, glutamate dehydrogenase. ylammonium bromide in the concentrations given above. The domain of cytochrome b5, the trypsin form of aminopeptidise, protein samples used for agarose electrophoresis and immu- and the bacterial exopenicillinase. Cytochrome b5 (15) and noelectrophoresis usually contained 0.7-1.5 mg of protein per membrane penicillinase (K. Simons and M. Sarvas, unpublished ml dissolved in the electrophoretic buffer containing a 4-fold results) consist of single polypeptide chains when solubilized detergent concentration over that used in the gels. The samples with Triton X-100, whereas the membrane aminopeptidase were allowed to equilibrate overnight at 00. contains three noncovalently bound polypeptide chains (16). Fig. 1 shows the electrophoretic patterns obtained for a RESULTS AND DISCUSSION number of ordinary soluble proteins in the absence of detergent We have tested 17 hydrophilic proteins and 5 amphiphilic and in the three detergent media. Only minor differences in The included acidic the electrophoretic mobility were observed. The same was true membrane proteins. hydrophilic proteins for exopenicillinase (Fig. 2), the trypsin form of aminopeptidase and basic proteins and glycoproteins, and some contained several subunits. Of the amphiphilic membrane proteins tested, (Fig. 2), and the polar fragment of cytochrome b5 (Fig. 3). In contrast, all the amphiphilic membrane proteins tested, three could be obtained in a proteolytically cleaved form that contained only the polar moiety: the polar heme-containing membrane penicillinase (Fig. 2), membrane aminopeptidase IIANE EXOPENICIWNASE PENKMLWHASE - ORIGIN 1 TX-DOC TX TX-CTAB TX-DOC TX TX-CTAB ~~~~~~d E TX C ORIGIN 2 I TX-CTAB AMINOPEPTIDASE MEMERANE (TRYPSIN FORM) AMWOPEPTIDASE 1cm FIG. 2. Combined results from agarose gel electrophoresis FIG. 3. Agarose electrophoresis of a mixture of cytochrome b5 (the of exopenicillinase, membrane penicillinase, the trypsin form of d-form) and trypsin-cleaved cytochrome b5 (the t-form) in Triton aminopeptidase, and membrane awminopeptidase in Triton X-100 and X-100 and sodium deoxycholate (TX-DOC), in Triton X-100 alone sodium deoxycholate (TX-DOC), in Triton X-100 alone (TX), and (TX), and in Triton X-100 and cetyltrimethylammonium bromide in Triton X-100 and cetyltrimethylammonium bromide (TX-GTAB). (TX-CTAB). Electrophoresis was performed for 75 min at 4.5 V/cm. Protein stain: Coomassie blue. The plates were dried and scanned at 415 and 450 nm. Downloaded by guest on October 2, 2021 Biochemistry:BProc.Helenius and Simons Natl. Acad. Sci. USA 74 (1977) 531 MEMBRANE PENICILLINASE X.I 0 EXO- ORIGIN -> I I "!9 PENICILLINASE MEMBRANE PENICILLINASE TX-DOC TX TX-CTAB EXO- FIG. 4. Combined results from agarose electrophoresis of [35Slmethionine-labeled spike glycoprotein El from Semliki Forest PENICILLINASE _ virus in the presence ofTriton X-100 and deoxycholate (TX-DOG), Triton X-100 alone (TX), and Triton X-100 and cetyltrimethylam- MEMBRANE monium bromide (TX-CTAB). The samples contained 8000 cpm in PENICILLINASE 12 Mtl (specific activity 4500 cpm/jug of protein). Electrophoresis was performed for 4 hr at 4.5 V/cm. After drying, the gels were autoradi- ographed for 6 days. EXO - PENICILLINASE _ FIG. 5. Immunoelectrophoresis of exopenicillinase and mem- brane penicillinase in the presence of Triton X-100 and sodium (Fig.
Load more

Recommended publications
  • Sodium Dodecyl Sulfate
    Catalog Number: 102918, 190522, 194831, 198957, 811030, 811032, 811033, 811034, 811036 Sodium dodecyl sulfate Structure: Molecular Formula: C12H25NaSO4 Molecular Weight: 288.38 CAS #: 151-21-3 Synonyms: SDS; Lauryl sulfate sodium salt; Dodecyl sulfate sodium salt; Dodecyl sodium sulfate; Sodium lauryl sulfate; Sulfuric acid monododecyl ester sodium salt Physical Appearance: White granular powder Critical Micelle Concentration (CMC): 8.27 mM (Detergents with high CMC values are generally easy to remove by dilution; detergents with low CMC values are advantageous for separations on the basis of molecular weight. As a general rule, detergents should be used at their CMC and at a detergent-to-protein weight ratio of approximately ten. 13,14 Aggregation Number: 62 Solubility: Soluble in water (200 mg/ml - clear, faint yellow solution), and ethanol (0.1g/10 ml) Description: An anionic detergent3 typically used to solubilize8 and denature proteins for electrophoresis.4,5 SDS has also been used in large-scale phenol extraction of RNA to promote the dissociation of protein from nucleic acids when extracting from biological material.12 Most proteins bind SDS in a ratio of 1.4 grams SDS to 1 gram protein. The charges intrinsic to the protein become insignificant compared to the overall negative charge provided by the bound SDS. The charge to mass ratio is essentially the same for each protein and will migrate in the gel based only on protein size. Typical Working Concentration: > 10 mg SDS/mg protein Typical Buffer Compositions: SDS Electrophoresis
    [Show full text]
  • Tutorial on Working with Micelles and Other Model Membranes
    Tutorial on Working with Micelles and Model Membranes Chuck Sanders Dept. of Biochemistry, Dept. of Medicine, and Center for Structural Biology Vanderbilt University School of Medicine. http://structbio.vanderbilt.edu/sanders/ March, 2017 There are two general classes of membrane proteins. This presentation is on working with integral MPs, which traditionally could be removed from the membrane only by dissolving the membrane with detergents or organic solvents. Multilamellar Vesicles: onion-like assemblies. Each layer is one bilayer. A thin layer of water separates each bilayer. MLVs are what form when lipid powders are dispersed in water. They form spontaneously. Cryo-EM Micrograph of a Multilamellar Vesicle (K. Mittendorf, C. Sanders, and M. Ohi) Unilamellar Multilamellar Vesicle Vesicle Advances in Anesthesia 32(1):133-147 · 2014 Energy from sonication, physical manipulation (such as extrusion by forcing MLV dispersions through filters with fixed pore sizes), or some other high energy mechanism is required to convert multilayered bilayer assemblies into unilamellar vesicles. If the MLVs contain a membrane protein then you should worry about whether the protein will survive these procedures in folded and functional form. Vesicles can also be prepared by dissolving lipids using detergents and then removing the detergent using BioBeads-SM dialysis, size exclusion chromatography or by diluting the solution to below the detergent’s critical micelle concentration. These are much gentler methods that a membrane protein may well survive with intact structure and function. From: Avanti Polar Lipids Catalog Bilayers can undergo phase transitions at a critical temperature, Tm. Native bilayers are usually in the fluid (liquid crystalline) phase.
    [Show full text]
  • Cleaning and Sanitizing I. Cleaning 2 Types Of
    Cleaning and Sanitizing CLEANING AND SANITIZING I. CLEANING 2 TYPES OF CLEANING COMPOUNDS 2 PROPERTIES OF A CLEANER 2 FACTORS THAT AFFECT CLEANING EFFICIENCY 2 CLEANING OPERATION 3 II. SANITIZING 4 HEAT 4 CHEMICAL SANITIZERS 5 FACTORS AFFECTING SANITIZING 7 III. DISHWASHING MACHINES 8 HOT WATER SANITIZING 8 CHEMICAL SANITIZING 9 REQUIREMENTS FOR A SUCCESSFUL DISHWASHING OPERATION 10 CHECKING A DISHMACHINE 10 COMMON PROBLEMS 12 IV. DEFINITIONS: 14 V. QUIZ 16 VI. ANSWER KEY TO QUIZ 18 VII. REFERENCES: 18 1 Cleaning and Sanitizing This section is presented primarily for information. The only information the BETC participant will be responsible for is to know the sanitization standards for chemical and hot water sanitizing as found in the Rules for Food Establishment Sanitation. CLEANING AND SANITIZING I. CLEANING Cleaning is a process which will remove soil and prevent accumulation of food residues which may decompose or support the growth of disease causing organisms or the production of toxins. Listed below are the five basic types of cleaning compounds and their major functions: 1. Basic Alkalis - Soften the water (by precipitation of the hardness ions), and saponify fats (the chemical reaction between an alkali and a fat in which soap is produced). 2. Complex Phosphates - Emulsify fats and oils, disperse and suspend oils, peptize proteins, soften water by sequestering, and provide rinsability characteristics without being corrosive. 3 Surfactant - (Wetting Agents) Emulsify fats, disperse fats, provide wetting properties, form suds, and provide rinsability characteristics without being corrosive. 4. Chelating - (Organic compounds) Soften the water by sequestering, prevent mineral deposits, and peptize proteins without being corrosive.
    [Show full text]
  • Linear Alkylbenzene Sulphonate (CAS No
    Environmental Risk Assessment LAS Linear Alkylbenzene Sulphonate (CAS No. 68411-30-3) Revised ENVIRONMENTAL Aspect of the HERA Report = February 2013 = 1 1. Contents 2. Executive summary 3. Substance characterisation 3.1 CAS No. and grouping information 3.2 Chemical structure and composition 3.3 Manufacturing route and production/volume statistics 3.4 Consumption scenario in Europe 3.5 Use application summary 4. Environmental safety assessment 4.1 Environmental exposure assessment 4.1.1 Biotic and abiotic degradability 4.1.2 Removal 4.1.3 Monitoring studies 4.1.4 Exposure assessment: scenario description 4.1.5 Substance data used for the exposure calculation 4.1.6 PEC calculations 4.1.7 Bioconcentration 4.2 Environmental effects assessment 4.2.1 Ecotoxicity 4.2.1.1 Aquatic ecotoxicity 4.2.1.2 Terrestrial ecotoxicity 4.2.1.3 Sediment ecotoxicity 4.2.1.4 Ecotoxicity to sewage microorganisms 4.2.1.5 Reassurance on absence of estrogenic effects 4.2.2 PNEC calculations 4.2.2.1 Aquatic PNEC 4.2.2.2 Terrestrial PNEC 4.2.2.3 Sludge PNEC 4.2.2.4 Sediment PNEC 4.2.2.5 STP PNEC 4.3 Environment risk assessment 5. 5. References 6. Contributors to the report 6.1 Substance team 6.2 HERA environmental task force 6.3 HERA human health task force 6.4 Industry coalition for the OECD/ICCA SIDS assessment of LAS 2 2. Executive Summary Linear alkylbenzene sulphonate (LAS) is an anionic surfactant. It was introduced in 1964 as the readily biodegradable replacement for highly branched alkylbenzene sulphonates (ABS).
    [Show full text]
  • Quaternary Ammonium Compositions and Their Uses
    Europaisches Patentamt (19) European Patent Office Office europeen des brevets (11) EP 0 726 246 A1 (12) EUROPEAN PATENT APPLICATION (43) Date of publication: (51) |nt. CI.6: C07C 21 1/63, C01 B 33/44, 14.08.1996 Bulletin 1996/33 C1 p 1/62j Q21 C 5/02, (21) Application number: 96101900.7 A61 K 7/50 //C09D7/12 (22) Date of filing: 09.02.1996 (84) Designated Contracting States: • Campbell, Barbara DE DK ES FR GB IT NL Bristol, PA 1 9007 (US) • Chiavoni, Araxi (30) Priority: 10.02.1995 US 385295 Trenton, N J 0861 0 (US) • Magauran, Edward (71 ) Applicant: RHEOX INTERNATIONAL, INC. Westhampton, NJ 08060 (US) Hightstown, New Jersey 08520 (US) (74) Representative: Strehl Schubel-Hopf Groening & (72) Inventors: Partner • Cody, Charles, Dr. Maximilianstrasse 54 Robbinsville, NJ 08691 (US) 80533 Munchen (DE) (54) Quaternary ammonium compositions and their uses (57) Quaternary ammonium compositions are described which are made using diluents including soya bean oil, caster oil, mineral oils, isoparaffin/naphthenic and coconut oil. Such diluents remain as diluents in the final product and generally have a vapor pressure of 1mm of Hg or less at 25°C, and are liquid at ambient temperature. The quaternary/ammonium diluent com- positions have low volatile organic compound emission rates and high flash points, and can be tailored to partic- ular applications. Such applications include use a fabric softeners, cosmetics ingredients, deinking additives, surfactants, and reaction materials in the manufacture of organoclays. < CO CM CO CM o Q_ LU Printed by Rank Xerox (UK) Business Services 2.13.0/3.4 EP 0 726 246 A1 Description BACKGROUND OF THE INVENTION 5 1 .
    [Show full text]
  • Soaps and Detergent Book
    ,â\ soAPS hù \-.-.1' ¿'>/'--'u\ r *gg, Ç DETERGENTS ,' '.-"- iI ' \' /'l'- ''t "*-**'*o'*q-å- þr,-'- COIUTEIUTS , Cleaning products play an essential role in our daily lives. By safely and effectively removing soils, germs HISTORY .........4 and other contaminants, they help us to stay healthy, I care for our homes and possessions, and make our ì surroundings more pleasant. The Soap and Detergent Association (SDA) recognizes that public understanding of the safety and benefits of cleaning products is critical to their proper use. So we've revised Soaþs and Detergents to feature the most current information in an easy-to-read format. This second edition summarizes key developments in the history of cleaning products; the science of how they work; the procedures used to evaluate their safety for people and the environment; the functions of various products and their ingredients; and the most common manufacturing processes. SDA hopes that consumers, educators, students, media, government officials, businesses and others ñnd Soaps and Detergents avaluable resource of information about cleaning products. o ' \o.-. t ¡: 2nd Edition li¡: orpq¿ The Soap ancl Detergent Association I :. I The Soap an4\Detergent Associatidn ^T',-¡ (\ "i' ) t'l T/ j *.'Ò. t., ,,./-t"'\\\,1 ,,! -\. r'í-\ HISTORY r The earlv Greeks bathed for Ð aesthetié reasons and apparently did not use soap. Instead, they I The origins of personal cleanliness cleaned their bodies with blocks of I date back to prehistoric times. Since clay, sand, pumice and ashes, then water is essential for life, the eadiest anointed themselves with oil, and people lived near water and knew scraped off the oil and dirt with a something about its cleansing 11 Records show that ancient metal instrument known as a strigil.
    [Show full text]
  • Surfactants, Soaps, and Detergents
    Surfactants, soaps, and detergents Case study # 3 Presented by Kimberly Quant and Maryon P. Strugstad October 1st, 2010 Materials included in reading package: 1. Manahan, S., E. (2011). Water chemistry. CRC Press. Tylor and Francis Group. Boka Taton, FL, US 2. Bunce, N. (1994) Environmental Chemistry. 2nd Ed. Wuerz Publishing Ltd. Winnipeg, Canada. 3. Sodium Laureth sulfate and SLS (01 April, 2003). National Industrial Chemicals Notification and Assessment Scheme (NICNAS). http://www.dweckdata.com/Research_files/SLS_compendium.pdf 4. Westgate, T. (17 Aug 2006). Switchable surfactants give on-demand emulsions. Royal Society of Chemistry. http://www.rsc.org/chemistryworld/News/2006/August/17080602.asp 5. Matthew, J.S. (3 Aug 2000). The biodegradation of surfactants in the environment. 6. J. A. Perales, M. A. Linear Alkylbenzene Sulphonates: Biodegradability and lsomeric Composition. Bull. Environ. Contam. Toxicol. (1999) 63:94-100. Springer-Verlag New York Inc. Further reading: 1. Tadros, T., F (2005). Applied surfactants: principles and applications. Wiley-VCH. 2.Cosmetic ingredient review (1983). Final Report on the Safety Assessment of Sodium Lauryl Sulfte and Ammonium Lauryl Sulfate. International Journal of Toxicology, Vol 2, No. 7, p. 127-181 3. Y, Liu et. al. (2006). Switchable surfactants. Science, Vol 313, p. 958-960 4. Coghlan, A. (17 August 2006). Oil and water mix and un-mix on demand. http://www.newscientist.com/article/dn9781 5. Staples, C., J et. al. (2001). Ultimate biodegradation of alkylphenol ethoxylate surfactants and their biodegradation intermediates. Environmental Toxicology and Chemistry, Vol. 20, No. 11, pp. 2450–2455 Case Study # 3 – Surfactants, soaps and detergents 1 Saponification – production of natural soap Cross-section of a micelle Case Study # 3 – Surfactants, soaps and detergents 2 Table 1.
    [Show full text]
  • Classification of Cleaning Agents Cleaning Agents Are Classified According to the Principle Method by Which Soil Or Stains Are Removed from the Surface
    Classification of Cleaning Agents Cleaning agents are classified according to the principle method by which soil or stains are removed from the surface. This will be determined by their composition. The principle classes are: • Water • Detergents • Abrasives • Degreasers • Acid cleaners • Organic solvents • Other cleaning agents 1. WATER: Water is the simplest cleaning agent and some form of dirt will be dissolved by it, but normally it is a poor cleaning agent if used alone. It becomes effective only if used in conjunction with some other agent, e.g. a detergent. Water serves to: • Carry the cleaning materials to the soil • Suspend the soil • Remove the suspended soil from the cleaning site • Rinse the detergent solution from the surface • Water has poor power of detergency because: • It has high surface tension and forms droplets • It has little wetting power • It is repelled by oil and grease • If shaken within oil the emulsion does not prevent the formation of large droplets • It has low surfactant effect (surface active agent) 2. DETERGENT: Detergents are those cleaning agents, which contain significant quantities of a group of chemicals known as ‘Surfactants’ (chemicals that have water and soil attracting properties). A number of other chemicals are frequently included to produce detergents suitable for a specific use. A good detergent should – • Reduce the surface tension of water so that the cleaning solution can penetrate the soil • Emulsify soil and lift it from the surface • Be soluble in cold water • Be effective in hard water and a wide range of temperatures. • Be hard on the surface that has to be cleaned.
    [Show full text]
  • Reconstitution of Membrane Proteins Into Model Membranes: Seeking Better Ways to Retain Protein Activities
    Int. J. Mol. Sci. 2013, 14, 1589-1607; doi:10.3390/ijms14011589 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Review Reconstitution of Membrane Proteins into Model Membranes: Seeking Better Ways to Retain Protein Activities Hsin-Hui Shen 1,2,*, Trevor Lithgow 1 and Lisandra L. Martin 2 1 Department of Biochemistry and Molecular Biology, Monash University, Melbourne 3800, Australia; E-Mail: [email protected] 2 School of Chemistry, Monash University, Clayton, VIC 3800, Australia; E-Mail: [email protected] * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +61-3-9545-8159. Received: 20 December 2012; in revised form: 9 January 2013 / Accepted: 10 January 2013 / Published: 14 January 2013 Abstract: The function of any given biological membrane is determined largely by the specific set of integral membrane proteins embedded in it, and the peripheral membrane proteins attached to the membrane surface. The activity of these proteins, in turn, can be modulated by the phospholipid composition of the membrane. The reconstitution of membrane proteins into a model membrane allows investigation of individual features and activities of a given cell membrane component. However, the activity of membrane proteins is often difficult to sustain following reconstitution, since the composition of the model phospholipid bilayer differs from that of the native cell membrane. This review will discuss the reconstitution of membrane protein activities in four different types of model membrane—monolayers, supported lipid bilayers, liposomes and nanodiscs, comparing their advantages in membrane protein reconstitution. Variation in the surrounding model environments for these four different types of membrane layer can affect the three-dimensional structure of reconstituted proteins and may possibly lead to loss of the proteins activity.
    [Show full text]
  • The Effect of Antibacterial Agents Triclosan and N-Alkyl on E. Coli Viability
    Western Washington University Western CEDAR WWU Honors Program Senior Projects WWU Graduate and Undergraduate Scholarship Fall 2000 The Effect of Antibacterial Agents Triclosan and N-alkyl on E. coli Viability Heidi Nielsen Western Washington University Follow this and additional works at: https://cedar.wwu.edu/wwu_honors Part of the Biology Commons Recommended Citation Nielsen, Heidi, "The Effect of Antibacterial Agents Triclosan and N-alkyl on E. coli Viability" (2000). WWU Honors Program Senior Projects. 328. https://cedar.wwu.edu/wwu_honors/328 This Project is brought to you for free and open access by the WWU Graduate and Undergraduate Scholarship at Western CEDAR. It has been accepted for inclusion in WWU Honors Program Senior Projects by an authorized administrator of Western CEDAR. For more information, please contact [email protected]. The Effect of Antibacterial Agents Triclosan and N-alkyl on E. coli iabli!Y Heidi Nielsen Advisor: Jeff Young, Biology Fall 2000 An equal opportunity university Honors Program Bellingham, Washington 98225-9089 (360)650-3034 Fax (360) 650-7305 HONORS THESIS In presenting this honors paper in partial requirements for a bachelor's degree at Western Washington University, I agree that the Library shall make its copies freely available for inspection. I further agree that extensive copying of this thesis is allowable only for scholarly purposes. It is understood that any publication of this thesis for commercial purposes or for financial gain shall not be allowed without my written permission. Signature Date I\ /15 )oa r I Heidi Nielsen Student ID WOOOS9898 Honors Program Senior Project Fall 2000 The Effect of Antibacterial Agents Triclosan and N-Alkyl on E.
    [Show full text]
  • Rigid Amphiphiles for Membrane Protein Manipulation
    COMMUNICATIONS [4] a) A. V. Eliseev, M. I. Nelen, J. Am. Chem. Soc. 1997, 119, 1147 ± 1148; Rigid Amphiphiles for Membrane Protein b) A. V. Eliseev, M. I. Nelen, Chem. Eur. J. 1998, 4, 825 ± 834. [5] a) P. A. Brady, J. K. M. Sanders, J. Chem. Soc. Perkin Trans. 1 1997, Manipulation** 3237 ± 3253; b) H. Hioki, W. C. Still, J. Org. Chem. 1998, 63, 904 ± 905; D. Tyler McQuade, Mariah A. Quinn, Seungju M. Yu, c) M. Albrecht, O. Bau, R. Fröhlich, Chem. Eur. J. 1999, 5, 48 ± 56; d) I. Huc, M. J. Krische, D. P. Funeriu, J.-M. Lehn, Eur. J. Inorg. Chem. Arthur S. Polans, Mark P. Krebs,* and 1999, 1415 ± 1420. Samuel H. Gellman* [6] For recent overviews, see: a) A. Ganesan, Angew. Chem. 1998, 110, 2989 ± 2992; Angew. Chem. Int. Ed. 1998, 37, 2828 ± 2831, and The shape of an amphiphile strongly influences self- references therein; b) C. Gennari, H. P. Nestler, U. Piarulli, B. Salom, association in solution[1] and in liquid crystalline phases,[2] as Liebigs Ann. 1997, 637 ± 647, and references therein; c) J.-M. Lehn, Chem. Eur. J. 1999, 5, 2455 ± 2463. well as interactions with self-assembled structures such as [7] Several examples of host-templated selection of the strongest binder lipid bilayers.[3] In recent years several groups have examined in a dynamic library of guest molecules were recently reported: a) I. unusual amphiphile topologies.[4, 5] We and others, for exam- Huc, J.-M. Lehn, Proc. Natl. Acad. Sci. USA 1997, 94, 2106 ± 2110; b) S.
    [Show full text]
  • Lipid-Detergent Phase Transitions During Detergent-Mediated Liposome Solubilization
    J Membrane Biol DOI 10.1007/s00232-016-9894-1 Lipid-Detergent Phase Transitions During Detergent-Mediated Liposome Solubilization 1,2 3 3 Hanieh Niroomand • Guru A. Venkatesan • Stephen A. Sarles • 1,2,3 1,2,3 Dibyendu Mukherjee • Bamin Khomami Received: 29 October 2015 / Accepted: 24 March 2016 Ó Springer Science+Business Media New York 2016 Abstract We investigate the phase transition stages for proteoliposome formation. Finally, although the solubi- detergent-mediated liposome solubilization of bio-mimetic lization of DPPG with DDM indicated the familiar three- membranes with the motivation of integrating membrane- stage process, the same process with TX-100 indicate bound Photosystem I into bio-hybrid opto-electronic structural deformation of vesicles into complex network of devices. To this end, the interaction of two non-ionic kinetically trapped micro- and nanostructured arrange- detergents n-dodecyl-b-D-maltoside (DDM) and Triton ments of lipid bilayers. X-100 (TX-100) with two types of phospholipids, namely DPhPC (1,2-diphytanoyl-sn-glycero-3-phosphocholine) Keywords Detergent-mediated liposome solubilization Á and DPPG (1,2-dipalmitoyl-sn-glycero-3-phospho-(10-rac- DPhPC and DPPG Á n-dodecyl-b-D-maltoside (DDM) and glycerol)), are examined. Specifically, solubilization pro- Triton X-100 (TX-100) Á Lamellar-to-micellar transition Á cesses for large unilamellar liposomes are studied with the Lamellar-to-complex structures Á Transmission electron aid of turbidity measurements, dynamic light scattering, microscopy and cryo-transmission electron microscopy imaging. Our results indicate that the solubilization process is well depicted by a three-stage model, wherein the lamellar-to- Introduction micellar transitions for DPhPC liposomes are dictated by the critical detergent/phospholipid ratios.
    [Show full text]