title: Proteomic profiling and identification of cofilin responding to oxidative stress in vascular smooth muscle.
abstract: We used 2-DE and MALDI-TOF/TOF to identify proteins of vascular smooth muscle cells whose expression was or was not altered by exposure to 500 microM H2O2 for 30 min. We detected more than 800 proteins on silver-stained gels of whole protein extracts from rat aortic smooth muscle strips. Of these proteins, 135 clearly unaffected and 19 having levels altered by exposure to H2O2 were identified. Protein characterization revealed that the most prominent vascular smooth muscle proteins were those with antioxidant, cytoskeletal structure, or muscle contraction. In addition, cofilin, an isoform of the actin depolymerizing factor family, shifted to its basic site on the 2-DE gel as a result of H2O2 treatment. In Western blot analysis of proteins from A7r5 aortic smooth muscle cells, the phosphorylation, but not the expression, of cofilin was decreased by H2O2 in a dose-dependent manner. The H2O2-induced dephosphorylation of cofilin and apoptosis was inhibited by Na3VO4, an inhibitor of protein tyrosine phosphatase (PTP). These results suggest that cofilin is one of the proteins regulated by H2O2 treatment in vascular smooth muscle, and has an important role in the induction of vascular apoptosis through PTP-dependent mechanisms.
authors.affiliation: Department of Physiology, Konkuk University, Danwol-dong, Chungju City, Chungbuk, Korea.
authors.forename: Chang-Kwon
authors.initials: CK
authors.lastname: Lee
authors.forename: Hyo-Jun
authors.initials: HJ
authors.lastname: Park
authors.forename: Hyeon Ha
authors.initials: HH
authors.lastname: So
authors.forename: Hyo Jin
authors.initials: HJ
authors.lastname: Kim
authors.forename: Keun Sang
authors.initials: KS
authors.lastname: Lee
authors.forename: Wahn Soo
authors.initials: WS
authors.lastname: Choi
authors.forename: Hwan Myung
authors.initials: HM
authors.lastname: Lee
authors.forename: Kyung-Jong
authors.initials: KJ
authors.lastname: Won
authors.forename: Taek Joon
authors.initials: TJ
authors.lastname: Yoon
authors.forename: Tae-Kyu
authors.initials: TK
authors.lastname: Park
authors.forename: Bokyung
authors.initials: B
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002460:Cell Line; D051340:Cofilin 2 / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D040081:Protein Array Analysis / Q000379:methods*; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014638:Vanadates / Q000494:pharmacology
chemical_list: D051340:Cofilin 2; D004791:Enzyme Inhibitors; D009124:Muscle Proteins; D014638:Vanadates; D006861:Hydrogen Peroxide; D017027:Protein Tyrosine Phosphatases