DOCSLIB.ORG

Mechanism–Based Inhibitors for Copper Amine Oxidases

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mechanism–Based Inhibitors for Copper Amine Oxidases MECHANISM–BASED INHIBITORS FOR COPPER AMINE OXIDASES: SYNTHESIS, MECHANISM, AND ENZYMOLOGY By BO ZHONG Submitted in partial fulfillment of the requirements For the degree of Doctor of Philosophy Thesis Adviser: Dr. Lawrence M. Sayre, Dr Irene Lee Department of Chemistry CASE WESTERN RESERVE UNIVERSITY January, 2010 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the thesis/dissertation of ______________________________________________________ candidate for the ________________________________degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. Table of Contents Table of Contents ............................................................................................................... I List of Tables .................................................................................................................... V List of Figures .................................................................................................................. VI List of Schemes ................................................................................................................ XI Acknowledgements ...................................................................................................... XIV List of Abbreviations ..................................................................................................... XV Abstract ........................................................................................................................ XVII Chapter 1. Introduction ................................................................................................... 1 1.1 General introduction to copper amine oxidases (CAOs) .......................................... 2 1.2 Mechanism for CAOs-catalyzed turnover................................................................. 3 1.3 General introduction to mechanism-based inhibitors................................................ 8 1.4 Kinetics for mechanism-based inhibition .................................................................. 9 1.5 References ............................................................................................................... 13 Chapter 2. Inactivation of two different copper amine oxidases by allenic amines 15 2.1 Introduction ............................................................................................................. 16 I 2.2 Results ..................................................................................................................... 18 2.2.1 Synthesis............................................................................................................... 18 2.2.2 Inactivation of bovine plasma amine oxidase by allenic amines ........................ 20 2.2.3 Inactivation of Arthrobacter globiformis amine oxidase (AGAO) by allenic amines............................................................................................................................ 27 2.2.4 Structure of AGAO-inhibitor rac-2 complex ....................................................... 29 2.3 Discussion and conclusion ...................................................................................... 30 2.4 Experimental procedures ......................................................................................... 38 2.5 References ............................................................................................................... 47 Chapter 3. Inactivation of bovine plasma amine oxidase (BPAO) by acetic acid-3- amino-propenyl ester ...................................................................................................... 49 3.1 Introduction ............................................................................................................. 50 3.2 Results and discussion ............................................................................................. 53 3.2.1 Synthesis............................................................................................................... 53 3.2.2 Enzyme assay of inhibition of BPAO by (E)- and (Z)-acetic acid-3-amino- propenyl ester (1 and 2) ................................................................................................. 54 3.2.3 Metabolism of acetic acid (E)-3-amino-propenyl ester (1) by BPAO ................. 56 3.2.4 Enzyme assay of inhibition of BPAO by aldehyde product 3-(acetyloxy)-(E)-2- propenal and its hydrolytic product MA ....................................................................... 59 II 3.2.5 Evaluation of 3-amino-propionaldehyde and its analogs as inhibitors and substrates of BPAO ....................................................................................................... 60 3.3 Mechanistic conclusion ........................................................................................... 72 3.4 Experimental procedures ......................................................................................... 76 3.5 References ............................................................................................................... 85 Chapter 4. Kinetic analysis of propargylamine derivatives as both substrates and inhibitors for bovine plasma amine oxidase (BPAO) and Arthrobacter globiformis amine oxidase (AGAO) ................................................................................................... 87 4.1 Introduction ............................................................................................................. 88 4.2 Results and discussion ............................................................................................. 92 4.2.1 Steady-state kinetic parameters for AGAO/BPAO-catalyzed oxidation of propargylamine derivatives ........................................................................................... 92 4.2.2 Kinetic parameters for inactivation of AGAO/BPAO by propargylamine derivatives ..................................................................................................................... 94 4.2.3 Inactivation of substrate-reduced BPAO by α, β-unsaturated aldehydes ............. 99 4.2.4 Anaerobic inactivation of BPAO by propargylamine derivatives ..................... 102 4.3 Conclusions ........................................................................................................... 104 4.4 Experimental procedures ....................................................................................... 105 4.5 References ............................................................................................................. 111 III Chapter 5. Investigate structural basis of inactivation of bovine plasma amine oxidase (BPAO) by 5-aryl-3-pentynamines ................................................................ 114 5.1 Introduction ........................................................................................................... 115 5.2 Results ................................................................................................................... 116 5.2.1 Synthesis............................................................................................................. 116 5.2.2 Irreversible inactivation of BPAO by 5-aryl-3-pentynamines (1-3) .................. 117 5.2.3 Mechanism of inactivation of BPAO by 5-aryl-3-pentynamines....................... 121 5.2.3.1 Covalent adduct between BPAO and 5-aryl-3-pentynamines......................... 121 5.2.3.2 Immunochemical analysis of inactivation of enzyme BPAO by inhibitor 3 .. 126 5.2.3.3 Attempt to resolve the structural basis of inactivation of BPAO by inhibitor 2 using enzymatic digestion and LC/MS ....................................................................... 127 5.3 Discussion ............................................................................................................. 130 5. 4 Conclusions .......................................................................................................... 134 5. 5 Experimental procedures ...................................................................................... 135 5.6 References ............................................................................................................. 144 Appendix ........................................................................................................................ 146 Bibliography .................................................................................................................. 152 IV List of Tables Chapter 2 ......................................................................................................................... 15 Table 2.1 The inhibitory potencies and reversibility of inhibition on BPAO by allene compounds ........................................................................................................................ 21 Table 2.2 The inhibitory potencies and reversibility of inhibition on AGAO by allene compounds ........................................................................................................................ 28 Chapter 4 ........................................................................................................................
Load more

Recommended publications
  • Zinc-Α2-Glycoprotein Is an Inhibitor of Amine Oxidase
    1 Supplemental Fig. 1 and 2 of “ZINC-α2-GLYCOPROTEIN IS AN INHIBITOR OF AMINE 2 OXIDASE COPPER-CONTAINING 3”, Matthias Romauch 3 Plasma enhances AOC3 activity 4 Endogenous plasma amine oxidase activity has previously been described in both human and 5 mouse plasma [1–3]. This activity derives from membrane-bound AOC3 which has been 6 released by metalloprotease activity [4]. A pronounced increase in levels of cleaved AOC3 is 7 observed during diabetes, congestive heart failure and liver cirrhosis [5–7]. Incubating 8 recombinant AOC3 with IEX fractions lacking ZAG (Fig. 4, B) increased amine oxidase 9 activity, which might be due to plasma-derived AOC3 activity or other plasma components. 10 To test this hypothesis, the amine oxidase activity in plasma of wt, AOC3 k.o. and ZAG k.o. 11 mice was measured directly using radioactive benzylamine as substrate. The activity linearly 12 increased with measured plasma volume of wt and ZAG k.o. mice, and the activity could be 13 blocked by the highly selective AOC3 inhibitor LJP1586 (Supplemental Fig. 1, A and B). 14 However, AOC3 activity in AOC3 k.o. plasma does not increase with increasing plasma 15 volume and residual activity cannot be further significantly reduced by adding LJP1586 16 (Supplemental Fig. 1, C). This suggests that AOC3 is the main plasma enzyme responsible for 17 benzylamine deamination, but it does not exclude other amine oxidases that are not sensitive 18 to LJP1586 or that have a higher affinity for other substrates. One such category of enzymes 19 could be the lysyl oxidases, which are also members of the copper amine oxidase family.
    [Show full text]
  • Topical LOX Inhibitor  2 Compounds Phase 2 Ready in 2020
    Investor Presentation Gary Phillips CEO 26 July 2019 For personal use only 1 Forward looking statement This document contains forward-looking statements, including statements concerning Pharmaxis’ future financial position, plans, and the potential of its products and product candidates, which are based on information and assumptions available to Pharmaxis as of the date of this document. Actual results, performance or achievements could be significantly different from those expressed in, or implied by, these forward-looking statements. All statements, other than statements of historical facts, are forward-looking statements. These forward-looking statements are not guarantees or predictions of future results, levels of performance, and involve known and unknown risks, uncertainties and other factors, many of which are beyond our control, and which may cause actual results to differ materially from those expressed in the statements contained in this document. For example, despite our efforts there is no certainty that we will be successful in partnering our LOXL2 program or any of the other products in our pipeline on commercially acceptable terms, in a timely fashion or at all. Except as required by law we undertake no obligation to update these forward-looking statements as a result of new information, future events or otherwise. For personal use only 2 A business model generating a valuable pipeline in fibrotic and inflammatory diseases • Anti fibrosis / oncology drug successfully cleared initial Clinical phase 1 study trials in high
    [Show full text]
  • Neurotransmitter Resource Guide
    NEUROTRANSMITTER RESOURCE GUIDE Science + Insight doctorsdata.com Doctor’s Data, Inc. Neurotransmitter RESOURCE GUIDE Table of Contents Sample Report Sample Report ........................................................................................................................................................................... 1 Analyte Considerations Phenylethylamine (B-phenylethylamine or PEA) ................................................................................................. 1 Tyrosine .......................................................................................................................................................................................... 3 Tyramine ........................................................................................................................................................................................4 Dopamine .....................................................................................................................................................................................6 3, 4-Dihydroxyphenylacetic Acid (DOPAC) ............................................................................................................... 7 3-Methoxytyramine (3-MT) ............................................................................................................................................... 9 Norepinephrine ........................................................................................................................................................................
    [Show full text]
  • Increased Vascular Adhesion Protein 1 (VAP-1) Levels Are Associated with Alternative M2 Macrophage Activation and Poor Prognosis for Human Gliomas
    diagnostics Article Increased Vascular Adhesion Protein 1 (VAP-1) Levels Are Associated with Alternative M2 Macrophage Activation and Poor Prognosis for Human Gliomas Shu-Jyuan Chang 1 , Hung-Pin Tu 2, Yen-Chang Clark Lai 3, Chi-Wen Luo 4,5, Takahide Nejo 6, 6 3,7,8, , 9,10, , Shota Tanaka , Chee-Yin Chai * y and Aij-Lie Kwan * y 1 Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] 2 Department of Public Health and Environmental Medicine, School of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; [email protected] 3 Department of Pathology, Kaohsiung Medical University Chung Ho Memorial Hospital, Kaohsiung 80756, Taiwan; [email protected] 4 Division of Breast Surgery, Department of Surgery, Kaohsiung Medical University Chung Ho Memorial Hospital, Kaohsiung 80756, Taiwan; [email protected] 5 Department of Surgery, Kaohsiung Medical University Chung Ho Memorial Hospital, Kaohsiung 80756, Taiwan 6 Department of Neurosurgery, Graduate School of Medicine, University of Tokyo, Tokyo 113-0033, Japan; [email protected] (T.N.); [email protected] (S.T.) 7 Department of Pathology, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan 8 Institute of Biomedical Sciences, National Sun Yat-Sen University, Kaohsiung 80424, Taiwan 9 Department of Neurosurgery, Kaohsiung Medical University Chung Ho Memorial Hospital, Kaohsiung 80756, Taiwan 10 Department of Surgery, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan * Correspondence: [email protected] (C.-Y.C.); [email protected] (A.-L.K.); Tel.: +88-6-7312-1101 (ext.
    [Show full text]
  • The Role of Protein Crystallography in Defining the Mechanisms of Biogenesis and Catalysis in Copper Amine Oxidase
    Int. J. Mol. Sci. 2012, 13, 5375-5405; doi:10.3390/ijms13055375 OPEN ACCESS International Journal of Molecular Sciences ISSN 1422-0067 www.mdpi.com/journal/ijms Review The Role of Protein Crystallography in Defining the Mechanisms of Biogenesis and Catalysis in Copper Amine Oxidase Valerie J. Klema and Carrie M. Wilmot * Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, 321 Church St. SE, Minneapolis, MN 55455, USA; E-Mail: [email protected] * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +1-612-624-2406; Fax: +1-612-624-5121. Received: 6 April 2012; in revised form: 22 April 2012 / Accepted: 26 April 2012 / Published: 3 May 2012 Abstract: Copper amine oxidases (CAOs) are a ubiquitous group of enzymes that catalyze the conversion of primary amines to aldehydes coupled to the reduction of O2 to H2O2. These enzymes utilize a wide range of substrates from methylamine to polypeptides. Changes in CAO activity are correlated with a variety of human diseases, including diabetes mellitus, Alzheimer’s disease, and inflammatory disorders. CAOs contain a cofactor, 2,4,5-trihydroxyphenylalanine quinone (TPQ), that is required for catalytic activity and synthesized through the post-translational modification of a tyrosine residue within the CAO polypeptide. TPQ generation is a self-processing event only requiring the addition of oxygen and Cu(II) to the apoCAO. Thus, the CAO active site supports two very different reactions: TPQ synthesis, and the two electron oxidation of primary amines. Crystal structures are available from bacterial through to human sources, and have given insight into substrate preference, stereospecificity, and structural changes during biogenesis and catalysis.
    [Show full text]
  • Effects of an Anti-Inflammatory VAP-1/SSAO Inhibitor, PXS-4728A
    Schilter et al. Respiratory Research (2015) 16:42 DOI 10.1186/s12931-015-0200-z RESEARCH Open Access Effects of an anti-inflammatory VAP-1/SSAO inhibitor, PXS-4728A, on pulmonary neutrophil migration Heidi C Schilter1*†, Adam Collison2†, Remo C Russo3,4†, Jonathan S Foot1, Tin T Yow1, Angelica T Vieira4, Livia D Tavares4, Joerg Mattes2, Mauro M Teixeira4 and Wolfgang Jarolimek1,5 Abstract Background and purpose: The persistent influx of neutrophils into the lung and subsequent tissue damage are characteristics of COPD, cystic fibrosis and acute lung inflammation. VAP-1/SSAO is an endothelial bound adhesion molecule with amine oxidase activity that is reported to be involved in neutrophil egress from the microvasculature during inflammation. This study explored the role of VAP-1/SSAO in neutrophilic lung mediated diseases and examined the therapeutic potential of the selective inhibitor PXS-4728A. Methods: Mice treated with PXS-4728A underwent intra-vital microscopy visualization of the cremaster muscle upon CXCL1/KC stimulation. LPS inflammation, Klebsiella pneumoniae infection, cecal ligation and puncture as well as rhinovirus exacerbated asthma models were also assessed using PXS-4728A. Results: Selective VAP-1/SSAO inhibition by PXS-4728A diminished leukocyte rolling and adherence induced by CXCL1/KC. Inhibition of VAP-1/SSAO also dampened the migration of neutrophils to the lungs in response to LPS, Klebsiella pneumoniae lung infection and CLP induced sepsis; whilst still allowing for normal neutrophil defense function, resulting in increased survival. The functional effects of this inhibition were demonstrated in the RV exacerbated asthma model, with a reduction in cellular infiltrate correlating with a reduction in airways hyperractivity.
    [Show full text]
  • Effects of the Inhibitor of Glutamate Decarboxylase on the Development
    RSC Advances View Article Online PAPER View Journal | View Issue Effects of the inhibitor of glutamate decarboxylase on the development and GABA accumulation in Cite this: RSC Adv.,2018,8,20456 germinating fava beans under hypoxia-NaCl stress Yongqi Yin, Chao Cheng and Weiming Fang* Glutamate decarboxylase (GAD) is the key enzyme in GABA shunt, which catalyzes the a-decarboxylation of glutamate to produce GABA. A specific inhibitor for GAD is convenient to study the dynamic balances of GABA metabolism in plants. The inhibitor of GAD in germinated fava beans was screened, and its inhibitory effect on the growth and GABA accumulation in fava beans during germination under hypoxia- NaCl stress was investigated. The inhibitory effect of aminoxyacetate for fava bean GAD was better than those of other chemicals, and it increased with the increase in concentration in vivo. After aminoxyacetate (5 mM) application for 4 days during germination, the GAD activity in germinating fava beans was significantly inhibited by more than 90% in both organs. Meanwhile, the growth of fava bean Creative Commons Attribution-NonCommercial 3.0 Unported Licence. sprouts was also slightly suppressed. Moreover, the GABA contents decreased by 43.9% and 81.5% in a 4 Received 8th May 2018 day-old cotyledon and embryo, respectively, under aminoxyacetate treatment compared with that in the Accepted 23rd May 2018 control. In summary, these results showed that aminoxyacetate can serve as a specific inhibitor of GAD DOI: 10.1039/c8ra03940b in plants. At least 43.9% and 81.5% of GABA in germinating fava beans under hypoxia-NaCl stress were rsc.li/rsc-advances synthesized via GABA shunt.
    [Show full text]
  • Supplementary Table S4. FGA Co-Expressed Gene List in LUAD
    Supplementary Table S4. FGA co-expressed gene list in LUAD tumors Symbol R Locus Description FGG 0.919 4q28 fibrinogen gamma chain FGL1 0.635 8p22 fibrinogen-like 1 SLC7A2 0.536 8p22 solute carrier family 7 (cationic amino acid transporter, y+ system), member 2 DUSP4 0.521 8p12-p11 dual specificity phosphatase 4 HAL 0.51 12q22-q24.1histidine ammonia-lyase PDE4D 0.499 5q12 phosphodiesterase 4D, cAMP-specific FURIN 0.497 15q26.1 furin (paired basic amino acid cleaving enzyme) CPS1 0.49 2q35 carbamoyl-phosphate synthase 1, mitochondrial TESC 0.478 12q24.22 tescalcin INHA 0.465 2q35 inhibin, alpha S100P 0.461 4p16 S100 calcium binding protein P VPS37A 0.447 8p22 vacuolar protein sorting 37 homolog A (S. cerevisiae) SLC16A14 0.447 2q36.3 solute carrier family 16, member 14 PPARGC1A 0.443 4p15.1 peroxisome proliferator-activated receptor gamma, coactivator 1 alpha SIK1 0.435 21q22.3 salt-inducible kinase 1 IRS2 0.434 13q34 insulin receptor substrate 2 RND1 0.433 12q12 Rho family GTPase 1 HGD 0.433 3q13.33 homogentisate 1,2-dioxygenase PTP4A1 0.432 6q12 protein tyrosine phosphatase type IVA, member 1 C8orf4 0.428 8p11.2 chromosome 8 open reading frame 4 DDC 0.427 7p12.2 dopa decarboxylase (aromatic L-amino acid decarboxylase) TACC2 0.427 10q26 transforming, acidic coiled-coil containing protein 2 MUC13 0.422 3q21.2 mucin 13, cell surface associated C5 0.412 9q33-q34 complement component 5 NR4A2 0.412 2q22-q23 nuclear receptor subfamily 4, group A, member 2 EYS 0.411 6q12 eyes shut homolog (Drosophila) GPX2 0.406 14q24.1 glutathione peroxidase
    [Show full text]
  • Termin Translat Trna Utr Mutat Protein Signal
    Drugs & Chemicals 1: Tumor Suppressor Protein p53 2: Heterogeneous-Nuclear Ribonucleo- (1029) proteins (14) activ apoptosi arf cell express function inactiv induc altern assai associ bind mdm2 mutat p53 p73 pathwai protein regul complex detect exon famili genom respons suppress suppressor tumor wild-typ interact intron isoform nuclear protein sensit site specif splice suggest variant 3: RNA, Transfer (110) 4: DNA Primers (1987) codon contain differ eukaryot gene initi amplifi analysi chain clone detect dna express mrna protein region ribosom rna fragment gene genotyp mutat pcr sequenc site speci suggest synthesi polymorph popul primer reaction region restrict sequenc speci termin translat trna utr 5: Saccharomyces cerevisiae Proteins 6: Apoptosis Regulatory Proteins (291) (733) activ apoptosi apoptosis-induc albican bud candida cerevisia complex encod apoptot bcl-2 caspas caspase-8 cell eukaryot fission function growth interact involv death fasl induc induct ligand methyl necrosi pathwai program sensit surviv trail mutant pomb protein requir saccharomyc strain suggest yeast 7: Plant Proteins (414) 8: Membrane Proteins (1608) access arabidopsi cultivar flower hybrid leaf leav apoptosi cell conserv domain express function gene human identifi inhibitor line maiz plant pollen rice root seed mammalian membran mice mous mutant seedl speci thaliana tomato transgen wheat mutat protein signal suggest transport 1 9: Tumor Suppressor Proteins (815) 10: 1-Phosphatidylinositol 3-Kinase activ arrest cell cycl cyclin damag delet dna (441) 3-kinas activ
    [Show full text]
  • Metabolism of 3-Nitrotyrosine Induces Apoptotic Death in Dopaminergic Cells
    6124 • The Journal of Neuroscience, June 7, 2006 • 26(23):6124–6130 Neurobiology of Disease Metabolism of 3-Nitrotyrosine Induces Apoptotic Death in Dopaminergic Cells Be´atrice Blanchard-Fillion,1 Delphine Prou,2 Manuela Polydoro,1 David Spielberg,1 Elpida Tsika,1 Zeneng Wang,5 Stanley L. Hazen,5 Michael Koval,6 Serge Przedborski,2,3,4 and Harry Ischiropoulos1,7 1Stokes Research Institute, Children’s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104, Departments of 2Neurology and 3Pathology and Cell Biology and 4Center of Neurobiology and Behavior, Columbia University, New York, New York 10032, 5Department of Cardiovascular Medicine and Center for Cardiovascular Diagnostics and Prevention, Cleveland Clinic Foundation, Cleveland, Ohio 44195, and Departments of 6Physiology and 7Pharmacology, University of Pennsylvania, Philadelphia, Pennsylvania 19104 Intrastriatal injection of 3-nitrotyrosine, which is a biomarker for nitrating oxidants, provokes dopaminergic neuronal death in rats by unknown mechanisms. Herein, we show that extracellular 3-nitrotyrosine is transported via the L-aromatic amino acid transporter in nondopaminergic NT2 cells, whereas in dopaminergic PC12 cells, it is transported by both the l-aromatic amino acid and the dopamine transporters. In both cell lines, 3-nitrotyrosine is a substrate for tyrosine tubulin ligase, resulting in its incorporation into the C terminus of ␣-tubulin. In NT2 cells, incorporation of 3-nitrotyrosine into ␣-tubulin induces a progressive, reversible reorganization of the micro- tubule architecture. In PC12 cells, 3-nitrotyrosine decreases intracellular dopamine levels and is metabolized by the concerted action of the aromatic amino acid decarboxylase and monoamine oxidase. Intracellular levels of 133 ␮mol of 3-nitrotyrosine per mole of tyrosine did not alter NT2 viability but induced PC12 apoptosis.
    [Show full text]
  • Downloaded 18 July 2014 with a 1% False Discovery Rate (FDR)
    UC Berkeley UC Berkeley Electronic Theses and Dissertations Title Chemical glycoproteomics for identification and discovery of glycoprotein alterations in human cancer Permalink https://escholarship.org/uc/item/0t47b9ws Author Spiciarich, David Publication Date 2017 Peer reviewed|Thesis/dissertation eScholarship.org Powered by the California Digital Library University of California Chemical glycoproteomics for identification and discovery of glycoprotein alterations in human cancer by David Spiciarich A dissertation submitted in partial satisfaction of the requirements for the degree Doctor of Philosophy in Chemistry in the Graduate Division of the University of California, Berkeley Committee in charge: Professor Carolyn R. Bertozzi, Co-Chair Professor David E. Wemmer, Co-Chair Professor Matthew B. Francis Professor Amy E. Herr Fall 2017 Chemical glycoproteomics for identification and discovery of glycoprotein alterations in human cancer © 2017 by David Spiciarich Abstract Chemical glycoproteomics for identification and discovery of glycoprotein alterations in human cancer by David Spiciarich Doctor of Philosophy in Chemistry University of California, Berkeley Professor Carolyn R. Bertozzi, Co-Chair Professor David E. Wemmer, Co-Chair Changes in glycosylation have long been appreciated to be part of the cancer phenotype; sialylated glycans are found at elevated levels on many types of cancer and have been implicated in disease progression. However, the specific glycoproteins that contribute to cell surface sialylation are not well characterized, specifically in bona fide human cancer. Metabolic and bioorthogonal labeling methods have previously enabled enrichment and identification of sialoglycoproteins from cultured cells and model organisms. The goal of this work was to develop technologies that can be used for detecting changes in glycoproteins in clinical models of human cancer.
    [Show full text]
  • Human AOC3 ELISA Kit (ARG81991)
    Product datasheet [email protected] ARG81991 Package: 96 wells Human AOC3 ELISA Kit Store at: 4°C Component Cat. No. Component Name Package Temp ARG81991-001 Antibody-coated 8 X 12 strips 4°C. Unused strips microplate should be sealed tightly in the air-tight pouch. ARG81991-002 Standard 2 X 50 ng/vial 4°C ARG81991-003 Standard/Sample 30 ml (Ready to use) 4°C diluent ARG81991-004 Antibody conjugate 1 vial (100 µl) 4°C concentrate (100X) ARG81991-005 Antibody diluent 12 ml (Ready to use) 4°C buffer ARG81991-006 HRP-Streptavidin 1 vial (100 µl) 4°C concentrate (100X) ARG81991-007 HRP-Streptavidin 12 ml (Ready to use) 4°C diluent buffer ARG81991-008 25X Wash buffer 20 ml 4°C ARG81991-009 TMB substrate 10 ml (Ready to use) 4°C (Protect from light) ARG81991-010 STOP solution 10 ml (Ready to use) 4°C ARG81991-011 Plate sealer 4 strips Room temperature Summary Product Description ARG81991 Human AOC3 ELISA Kit is an Enzyme Immunoassay kit for the quantification of Human AOC3 in serum, plasma (heparin, EDTA) and cell culture supernatants. Tested Reactivity Hu Tested Application ELISA Specificity There is no detectable cross-reactivity with other relevant proteins. Target Name AOC3 Conjugation HRP Conjugation Note Substrate: TMB and read at 450 nm. Sensitivity 0.39 ng/ml Sample Type Serum, plasma (heparin, EDTA) and cell culture supernatants. Standard Range 0.78 - 50 ng/ml Sample Volume 100 µl www.arigobio.com 1/3 Precision Intra-Assay CV: 5.8%; Inter-Assay CV: 6.6% Alternate Names HPAO; Semicarbazide-sensitive amine oxidase; VAP-1; Vascular adhesion protein 1; Copper amine oxidase; SSAO; Membrane primary amine oxidase; EC 1.4.3.21; VAP1 Application Instructions Assay Time ~ 5 hours Properties Form 96 well Storage instruction Store the kit at 2-8°C.
    [Show full text]