PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES

PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES

Bibliographic Details
Main Author: Zhou, Gang
Language:English
Published: Cleveland State University / OhioLINK 2012
Subjects:
Biochemistry
Biomedical Research
Cellular Biology
Chemistry
Medicine
Protein Purification
HPLC
Drug Metabolism
Enzyme
Aspirin
Platelets
Cardiovascular Disease
Personalized Medicine
Clinical Chemistry
Online Access:http://rave.ohiolink.edu/etdc/view?acc_num=csu1338916020
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spelling ndltd-OhioLink-oai-etd.ohiolink.edu-csu13389160202021-08-03T05:35:19Z PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES Zhou, Gang Biochemistry Biomedical Research Cellular Biology Chemistry Medicine Protein Purification HPLC Drug Metabolism Enzyme Aspirin Platelets Cardiovascular Disease Personalized Medicine Clinical Chemistry Aspirin prophylaxis suppresses major adverse cardiovascular events, but its turnover in blood is rapid, which limits its inhibitory effects on platelet cyclooxygenase and thrombosis. Inter-individual variability of platelet responses to aspirin clinically presents as treatment failure, and this increasing clinical phenomenon is frequently named aspirin “resistance”. However, the molecular mechanisms behind this are unclear. Blood aspirin hydrolases are believed to control aspirin survival in vivo, but the identity of the circulating enzyme(s) that hydrolyzes aspirin remains unknown. In this thesis, blood aspirin hydrolases were identified and characterized. The relationship between blood aspirin hydrolases and aspirin efficacy was investigated. RP-HPLC analysis of the product salicylic acid with an internal standard showed plasma hydrolysis of aspirin varied 12-fold variation among 2,275 individuals. Genome-wide association analysis using serum aspirin hydrolytic activity from 2,275 individuals showed a genetic component to aspirin hydrolytic variation, and that only BChE significantly associated to aspirin variation. However, plasma from an individual with an inactivating point mutation in BChE effectively hydrolyzed aspirin. A non-BChE aspirin hydrolase was found in plasma, which can be distinguished from BChE by procainamide and oxidized ATP. Erythrocyte aspirin hydrolase was purified by 1400-fold and type I PAF acetylhydrolase was identified as a candidate aspirinase by mass spectrometry. Recombinant PAFAH1B2 hydrolyzed aspirin and aspirin was effectively hydrolyzed in cells ectopically expressing PAFAH1B2 and PAFAH1B3. Type I PAFAH also accounts for non-BChE plasma aspirin hydrolytic activity. Both plasma and erythrocytes were found modulated aspirin efficacy significantly, which varied by aspirin hydrolytic activity variation.I hope my work in this thesis will help to understand aspirin “resistance” and shed light on personalized medicine of aspirin. 2012-06-06 English text Cleveland State University / OhioLINK http://rave.ohiolink.edu/etdc/view?acc_num=csu1338916020 http://rave.ohiolink.edu/etdc/view?acc_num=csu1338916020 unrestricted This thesis or dissertation is protected by copyright: all rights reserved. It may not be copied or redistributed beyond the terms of applicable copyright laws.
collection NDLTD
language English
sources NDLTD
topic Biochemistry
Biomedical Research
Cellular Biology
Chemistry
Medicine
Protein Purification
HPLC
Drug Metabolism
Enzyme
Aspirin
Platelets
Cardiovascular Disease
Personalized Medicine
Clinical Chemistry
spellingShingle Biochemistry
Biomedical Research
Cellular Biology
Chemistry
Medicine
Protein Purification
HPLC
Drug Metabolism
Enzyme
Aspirin
Platelets
Cardiovascular Disease
Personalized Medicine
Clinical Chemistry
Zhou, Gang
PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES
author Zhou, Gang
author_facet Zhou, Gang
author_sort Zhou, Gang
title PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES
title_short PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES
title_full PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES
title_fullStr PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES
title_full_unstemmed PURIFICATION AND CHARACTERIZATION OF BLOOD ASPIRIN HYDROLASES
title_sort purification and characterization of blood aspirin hydrolases
publisher Cleveland State University / OhioLINK
publishDate 2012
url http://rave.ohiolink.edu/etdc/view?acc_num=csu1338916020
work_keys_str_mv AT zhougang purificationandcharacterizationofbloodaspirinhydrolases
_version_ 1719422177919369216

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