Identification of Cysteine, Methionine and Tryptophan Residues of Actin Oxidized In vivo during Oxidative Stress

被引:56
作者
Fedorova, Maria [1 ]
Kuleva, Nadezda [2 ]
Hoffmann, Ralf [1 ]
机构
[1] Univ Leipzig, Fac Chem & Mineral, Ctr Biotechnol & Biomed, Inst Bioanalyt Chem, D-04103 Leipzig, Germany
[2] St Petersburg State Univ, Fac Biol & Soil Sci, Dept Biochem, St Petersburg 199034, Russia
关键词
actin polymerization; ATPase activity of myosin; carbonylation; mass spectrometry (MS); protein oxidation; reactive oxygen species ROS; SKELETAL-MUSCLE FIBERS; MYOSIN ATPASE ACTIVITY; HYDROGEN-PEROXIDE; REACTIVE OXYGEN; FREE-RADICALS; FUNCTIONAL IMPAIRMENT; CONTRACTILE FUNCTION; NITROSATIVE STRESS; HEART-FAILURE; PROTEIN;
D O I
10.1021/pr901099e
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Increased levels of reactive oxygen species (ROS) cause oxidative stress and are believed to play a key role in the development of age-related diseases and mammalian aging in general by oxidizing proteins, lipids, and DNA. In this study, we have investigated the effects of ROS on actin in an established rat model of acute oxidative stress using short-term X-ray irradiation. Relative to the control, the actin functions studied in vitro were reduced for (i) actin polymerization to a minimum of 33% after 9 h and 00 actin activated Mg(2+)ATPase activity of myosin to 55% after 9 h. At 24 h, the activities had partially recovered to 64 and 80% of the control sample, respectively. The underlying oxidative modifications were also studied at the molecular level. The content of reactive carbonyl-groups increased 4-fold within the studied 24 h period. Among the five cysteine residues of actin, Cys(239) and Cys(259) were oxidized to sulfenic (Cys-SOH), sulfinic (CYS-SO2H), or suifonic (CYS-SO3H) acids by increasing amounts over the time periods studied. The content of methionine sulfoxides also increased for 15 of the 16 methionine residues, with Met(44), Met(47), and Met(355) having the highest sulfoxide contents. Met(82) was also further oxidized to the sulfone. Among the four tryptophan residues present in actin, only Trp(79) and Trp(86) appeared to undergo oxidation. The relative contents of hydroxy-tryptophan, N-formyl-kynurenine, and kynurenine increased after irradiation, reaching a maximum in the 9 h sample.
引用
收藏
页码:1598 / 1609
页数:12
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