Nitro-fatty Acid Reaction with Glutathione and Cysteine
Nitro-fatty Acid Reaction with Glutathione and Cysteine
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硝基脂肪酸与谷胱甘肽和半胱氨酸的反应
DOI：
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发表时间：
2007
影响因子：
4.8
通讯作者：
B. Freeman
中科院分区：
文献类型：
--
作者：
L. Baker;P. R. Baker;Franca Golin;F. Schopfer;Mitchell Fink;S. Woodcock;B. Branchaud;R. Radi;B. Freeman
由一氧化氮衍生的物种对脂肪酸的硝化产生亲电产物，这些产物与蛋白质硫醇加成，导致蛋白质功能和分布的变化。在健康人体血液中检测到蛋白质和还原型谷胱甘肽(GSH)的硝基脂肪酸加合物。动力学和质谱学分析表明，油酸(OA-NO2)和亚油酸(LNO2)的硝基烯烃衍生物通过Michael加成反应与GSH和Cys发生快速反应。停流分光光度法测得在pH 7.4和37℃时，OA-NO_2和LNO_2与GSH反应的二级速率常数分别为183m~(-1)s~(-1)和355m~(-1)s~(-1)，明显高于GSH与过氧化氢和非硝化亲电脂肪酸(包括8-异前列腺素A2和15-脱氧-Δ12，14-前列腺素J2)的反应速率。反应pH从7.4增加到8.9，硫醇与OA-NO2和LNO2反应的表观二级速率常数增大，表明反应活性依赖于GSH的硫酸盐阴离子。随着目标硫醇的pKa增加，硝基烯与硫醇的反应速率降低。增加洗涤剂辛基-β-d-吡喃葡萄糖苷的浓度会降低硝基烯与谷胱甘肽的反应速度，这表明硝基脂肪酸进入胶束或膜结构会限制Michael与更多极性亲核目标的反应。总之，这些结果揭示了硝基脂肪酸对硫醇的可逆加成是硝基脂肪酸对蛋白质功能进行可逆翻译后调节的一种机制。
Fatty acid nitration by nitric oxide-derived species yields electrophilic products that adduct protein thiols, inducing changes in protein function and distribution. Nitro-fatty acid adducts of protein and reduced glutathione (GSH) are detected in healthy human blood. Kinetic and mass spectrometric analyses reveal that nitroalkene derivatives of oleic acid (OA-NO2) and linoleic acid (LNO2) rapidly react with GSH and Cys via Michael addition reaction. Rates of OA-NO2 and LNO2 reaction with GSH, determined via stopped flow spectrophotometry, displayed second-order rate constants of 183 m-1s-1 and 355 m-1s-1, respectively, at pH 7.4 and 37 °C. These reaction rates are significantly greater than those for GSH reaction with hydrogen peroxide and non-nitrated electrophilic fatty acids including 8-iso-prostaglandin A2 and 15-deoxy-Δ12,14-prostaglandin J2. Increasing reaction pH from 7.4 to 8.9 enhanced apparent second-order rate constants for the thiol reaction with OA-NO2 and LNO2, showing dependence on the thiolate anion of GSH for reactivity. Rates of nitroalkene reaction with thiols decreased as the pKa of target thiols increased. Increasing concentrations of the detergent octyl-β-d-glucopyranoside decreased rates of nitroalkene reaction with GSH, indicating that the organization of nitro-fatty acids into micellar or membrane structures can limit Michael reactivity with more polar nucleophilic targets. In aggregate, these results reveal that the reversible adduction of thiols by nitro-fatty acids is a mechanism for reversible post-translational regulation of protein function by nitro-fatty acids.
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影响因子：
2.9
作者：
Han, D;Canali, R;Cadenas, E
通讯作者：
Cadenas, E
影响因子：
5.2
作者：
S. Woodcock;Adam J. V. Marwitz;P. Bruno;B. Branchaud
通讯作者：
S. Woodcock;Adam J. V. Marwitz;P. Bruno;B. Branchaud
DOI：
10.1073/pnas.92.19.8965
发表时间：
1995-09-12
影响因子：
11.1
作者：
PRESTERA, T;TALALAY, P
通讯作者：
TALALAY, P
DOI：
10.1073/pnas.85.21.8261
发表时间：
1988-11-01
影响因子：
11.1
作者：
TALALAY, P;DELONG, MJ;PROCHASKA, HJ
通讯作者：
PROCHASKA, HJ
影响因子：
5.2
作者：
Gorczynski, Michael J.;Huang, Jinming;King, S. Bruce
通讯作者：
King, S. Bruce