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Comparative Proteome Analysis of PAI‐1 and TNF‐α‐derived Endothelial Microparticles
Author(s) -
Ou Jingsong,
Kaul Sushma,
Greene Andrew S,
Pritchard Kirkwood A,
Oldham Keith T,
Sander Tara L
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a181-c
Subject(s) - tumor necrosis factor alpha , umbilical vein , proteome , chemistry , plasminogen activator , proteomics , plasminogen activator inhibitor 1 , tissue factor , microbiology and biotechnology , biology , biochemistry , in vitro , immunology , medicine , endocrinology , gene , coagulation
Endothelium‐derived microparticles (EMPs) are generated in response to cell injury, apoptosis, or activation via agonists such as plasminogen activator inhibitor type 1 (PAI‐1) and tumor necrosis factor alpha (TNF‐α). It is not known if different agonists produce EMPs with distinct biological functions. We hypothesize different agonists would generate EMPs with distinct protein composition. To test this hypothesis, EMPs were generated from PAI‐1 or TNF‐α‐stimulated human umbilical vein endothelial cells and subjected to protein analysis by two methods: 2‐D gel electrophoresis for subsequent proteomic analysis by MALD1‐TOF/MS and direct analysis by nano‐LC/MS analysis using FINNIGAN LTQ. We identified more than 60 proteins in both PAI‐1 and TNF‐α‐derived EMPs. PAI‐1 and TNF‐α derived EMPs are composed of overlapping, but distinct proteins that derive from different cellular components, exhibit multiple molecular functions, and are involved in various biological processes. Nano‐LC/MS analysis further confirmed the difference of EMPs profiles between PAI‐1 and TNF‐α stimulation. Our findings indicate that EMPs have overlapping, but potentially distinct protein composition dependent on the originating stimulus and provides fundamental insight into the mechanisms regulating the production of these particles and their physiologic role in different diseases.