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A practicable method to prepare nitrated proteins with peroxynitrite and low concentration of sodium hydroxide.
Molecular Biology Reports ( IF 2.8 ) Pub Date : 2019-11-30 , DOI: 10.1007/s11033-019-05211-w
Nan Zhang 1, 2, 3, 4 , Xiaochen Gao 1, 2, 3, 4 , Weijia Zhang 1, 2, 3, 4 , Doaa Higazy 1, 2, 3, 4 , Ke Wang 1, 2, 3, 4 , Zhenfang Fu 1, 2, 3, 4, 5 , Min Cui 1, 2, 3, 4
Affiliation  

Peroxynitrite is an ion acting as a powerful oxidant and nucleophile, which plays a key role in the inflammation and aging process by nitrating tyrosine or tryptophan residues of the proteins. Nitration of a target protein is considered to be a proper method to study the behavioral change of the proteins being nitrated. The commonly used methods for peroxynitrite preparation in vitro usually contain high concentration of sodium hydroxide, which easily induces hydrolysis of target proteins. Accordingly, the method for peroxynitrite preparation was optimized in vitro by changing the sequence of hydrochloric acid and sodium hydroxide added. After different amount of hydrochloric acid added to the system following sodium nitrite, peroxynitrite can be yielded in a concentration up to 60 mM with sodium hydroxide as low as 17 mM. More importantly, biological activity of the target protein was well maintained after protein nitration since low sodium hydroxide was used.

中文翻译:

用过氧亚硝酸盐和低浓度氢氧化钠制备硝化蛋白质的可行方法。

过氧亚硝酸盐是充当强大氧化剂和亲核试剂的离子,它通过硝化酪氨酸或蛋白质的色氨酸残基在炎症和衰老过程中发挥关键作用。靶蛋白的硝化被认为是研究被硝化的蛋白的行为变化的合适方法。体外过氧亚硝酸盐制备的常用方法通常包含高浓度的氢氧化钠,其容易诱导靶蛋白的水解。因此,通过改变加入盐酸和氢氧化钠的顺序,体外优化了过氧亚硝酸盐的制备方法。在亚硝酸钠后向系统中添加不同量的盐酸后,过氧化亚硝酸盐的浓度最高可达60 mM,而氢氧化钠的浓度低至17 mM。更重要的是,
更新日期:2019-11-01
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