Tyrosine nitration in human spermatozoa: a physiological function of peroxynitrite, the reaction product of nitric oxide and superoxide
Tyrosine nitration in human spermatozoa: a physiological function of peroxynitrite, the reaction product of nitric oxide and superoxide
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DOI：
10.1093/molehr/7.10.913
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发表时间：
2001-10-01
影响因子：
4
通讯作者：
Gagnon, C
中科院分区：
文献类型：
--
作者：
Herrero, MB;de Lamirande, E;Gagnon, C
酪氨酸硝化是一种广泛使用的过氧亚硝酸根（ONOO-）的标记物，其由一氧化氮（NO）反应产生。超氧阴离子（O-2（.-））。由于人类精子能够产生NO和O-2（.-）在体外获能过程中，我们研究了人精子中是否发生蛋白质的自发酪氨酸硝化，并评估了过氧亚硝酸盐对精子功能的生理效应。我们在这里报告，人类精子，孵育8小时的条件下，有利于获能，显示可重复的蛋白酪氨酸硝化模式。几种蛋白质与MOT。105-14 kDa的wt在15分钟孵育后变得越来越多地被酪氨酸硝化，然后观察到最小的变化。用人卵泡液或钙离子载体处理获能的精子，可引起运动时蛋白质中硝基酪氨酸含量的增加。wt为85 kDa。此外，精子暴露于ONOO-（2.5-50 μ mol/l）可增加精子活力，并使精子对人卵泡液的反应更早。ONOO-还以浓度依赖性方式增加蛋白质酪氨酸硝化和磷酸化。两者合计，这些结果表明，酪氨酸硝化的精子蛋白发生在获能的人精子，和低浓度的ONOO-调节精子功能，强调的概念，获能是氧化过程的一部分。
Tyrosine nitration is a widely used marker of peroxynitrite (ONOO-) produced from the reaction of nitric oxide (NO.) with superoxide (O-2(.-)). Since human spermatozoa are able to produce both NO. and O-2(.-) during capacitation in vitro, we investigated whether spontaneous tyrosine nitration of proteins occurs in human spermatozoa and evaluated the physiological effects of peroxynitrite on sperm function. We report here that human spermatozoa, incubated for 8 h under conditions conducive to capacitation, display a reproducible pattern of protein tyrosine nitration. Several proteins with mot. wt of 105-14 kDa become increasingly tyrosine-nitrated after 15 min incubation and then minimal changes are observed. Treatment of capacitated spermatozoa with human follicular fluid or calcium ionophore causes an increase of the nitrotyrosine content of proteins at the mot. wt of 85 kDa. Moreover, exposure of spermatozoa to ONOO- (2.5-50 mu mol/l) increases motility and primes spermatozoa to respond earlier to human follicular fluid. ONOO- also increases protein tyrosine nitration and phosphorylation in a concentration-dependent manner. Taken together, these results demonstrate that tyrosine nitration of sperm proteins occurs in capacitated human spermatozoa, and that low concentrations of ONOO- modulate sperm functions, emphasizing the concept that capacitation is part of an oxidative process.