Detection of protein adduction derived from styrene oxide to cysteine residues by alkaline permethylation.
Detection of protein adduction derived from styrene oxide to cysteine residues by alkaline permethylation.
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DOI：
10.1016/j.ab.2010.05.001
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发表时间：
2010-10-01
影响因子：
2.9
通讯作者：
Zheng J
中科院分区：
文献类型：
--
作者：
Dai J;Zhang F;Zheng J
氧化苯乙烯-半胱氨酸加合主要涉及蛋白质暴露于苯乙烯或氧化苯乙烯后在体内发生的共价修饰。在本研究中，我们开发了一种基于碱性过甲基化和GC/MS的方法来检测氧化苯乙烯衍生的蛋白加合。蛋白加合物的全甲基化产生两种甲硫基苯基乙醇，即2-甲硫基-2-苯基-1-乙醇和2-甲硫基-1-苯基-1-乙醇。为提高全甲基化效率，对反应温度、反应时间、NaOH浓度和CH 3 I/NaOH摩尔比等条件进行了研究。在优化条件下，半胱氨酸加合物的α和β异构体α-和β-巯基尿酸的全甲基化生成分析物的产率分别为35%和28%。氧化苯乙烯修饰的牛血清白蛋白的全甲基化释放出两种甲硫基苯乙醇，α-/β-加合比为1.5。在与不同浓度苯乙烯孵育的小鼠肝微粒体中观察到α-和β-加合的浓度依赖性增加。CD-1小鼠腹腔注射苯乙烯，剂量为0、50和400 mg/kg，每日一次，持续5天。在400 mg/kg组全血中观察到氧化苯乙烯衍生的蛋白加合物形成，α/β比为4.8，表明氧化苯乙烯与半胱氨酸残基的反应更可能发生在氧化苯乙烯的α-碳而不是β-碳上。
Styrene oxide-cysteine adduction predominantly involves in protein covalent modification undergoing in vivo after exposed to styrene or styrene oxide. In present study, we developed an alkaline permethylation- and GC/MS-based approach to detect styrene oxide-derived protein adduction. Permethylation of the protein adducts produced two methylthiophenylethanols, namely 2-methylthio-2-phenyl-1-ethanol and 2-methylthio-1-phenyl-1-ethanol. To improve the permethylation efficiency, reaction conditions, including temperature, time, NaOH strength and molar ratio of CH3I/NaOH, were explored. Under the optimized condition, the yields of the analyte formation resulting from permethylation of authentic standard α- and β-mercapturic acids, representing α and β isomers of cysteine adducts, were 35% and 28%, respectively. Permethylation of styrene oxide-modified bovine serum albumin released the two methylthiophenylethanols with an α-/β-adduction ratio of 1.5. A concentration-dependent increase in both α- and β-adduction was observed in mouse liver microsomes incubated with styrene at various concentrations. CD-1 mice were administered intraperitoneally with styrene at doses of 0, 50 and 400 mg/kg daily for 5 days. The formation of protein adducts derived from styrene oxide in whole blood in 400 mg/kg group was observed with an α/β ratio of 4.8, suggesting the reaction of styrene oxide with cysteine residues took place more likely at α-carbon than β-carbon of styrene oxide.
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影响因子：
5.9
作者：
Vodicka, Pavel;Koskinen, Mikko;Oesch, Franz
通讯作者：
Oesch, Franz
影响因子：
5.1
作者：
PACHECKA, J;GARIBOLDI, P;SALMONA, M
通讯作者：
SALMONA, M
影响因子：
4.1
作者：
RAPPAPORT, SM;TING, D;MCDONALD, T
通讯作者：
MCDONALD, T
影响因子：
4.7
作者：
HEMMINKI, K
通讯作者：
HEMMINKI, K
影响因子：
2.9
作者：
SLAUGHTER, DE;ZHENG, J;HANZLIK, RP
通讯作者：
HANZLIK, RP