硫化氢信号分子与<i>L</i>-乳酸脱氢酶蛋白相互作用机制的研究

引用本文: 朱艳雯, 刘玲, 孙玮洁, 颜廷才, 檀德宏, 张瑶, 白冰. 硫化氢信号分子与L-乳酸脱氢酶蛋白相互作用机制的研究[J]. 分析化学, 2018, 46(7): 1145-1151,1159. doi: 10.11895/j.issn.0253-3820.171510 shu

Citation: ZHU Yan-Wen, LIU Ling, SUN Wei-Jie, YAN Ting-Cai, TAN De-Hong, ZHANG Yao, Bai Bing. Investigation of Interaction Mechanism Between Signaling Molecule Hydrogen Sulfide and L-Lactic Dehydrogenase[J]. Chinese Journal of Analytical Chemistry, 2018, 46(7): 1145-1151,1159. doi: 10.11895/j.issn.0253-3820.171510 shu

硫化氢信号分子与L-乳酸脱氢酶蛋白相互作用机制的研究

朱艳雯 ^1, ,
刘玲 ^1, ,
孙玮洁 ^1, ,
颜廷才 ^1, ,
檀德宏 ^1, ,
张瑶 ^2, ,
白冰 ^1,

1.
沈阳农业大学食品学院, 沈阳 110866;
2.
沈阳师范大学美术与设计学院, 沈阳 110034
基金项目:
本文系国家自然科学基金项目（Nos.31571799，31571796）和十三五重点研发专项（No.2016YFD0400103）资助

摘要: 硫化氢（H₂S）作为信号分子与目标蛋白相互作用而实现信号传导机制的研究一直是研究热点。本研究以L-乳酸脱氢酶为目标蛋白，选用3种H₂S供体试剂（NaHS、Na₂S、PS），采用分子荧光实时监测L-乳酸脱氢酶活性变化，研究H₂S与目标蛋白的相互作用。SDS-PAGE电泳结果显示，H₂S不是通过形成二硫键/三硫键的方式与L-乳酸脱氢酶蛋白相互作用；圆二色谱结果表明，L-乳酸脱氢酶的二级结构发生变化，提示H₂S可能存在半胱氨酸巯基衍生能力，以硫烷硫存在的PS具有最强的巯基作用力；MALDI-TOF-MS/MS分析结果表明，L-乳酸脱氢酶蛋白中部分半胱氨酸巯基位点发生了硫烷化，进一步揭示了H₂S是通过与活性半胱氨酸巯基位点硫烷化的方式与L-乳酸脱氢酶发生作用。

关键词:

English

Investigation of Interaction Mechanism Between Signaling Molecule Hydrogen Sulfide and L-Lactic Dehydrogenase

1.
College of Food, Shenyang Agricultural University, Shenyang 110866, China;
2.
College of Fine Art and Design, Shenyang Normal University, Shenyang 110034, China
Received Date: 14 December 2017
Revised Date: 11 May 2018
Fund Project:
This work was supported by the National Natural Science Foundation of China (Nos. 31571799, 31571796) and the National Key Research Projects of China (No. 2016YFD0400103).

Abstract: Molecular mechanisms whereby H₂S influences its targets have been of intriguing interest. In this work, L-lactic dehydrogenase (L-LDH) was used as the protein target, and three kinds of H₂S-donor reagents (NaHS, Na₂S, and polysulfide) were chosen. The interactions of these H₂S-donor reagents with L-LDH were disclosed by molecular fluorescent assays for real-time monitoring of L-LDH activity. The results of the SDS-PAGE showed that H₂S might not interact with L-LDH to form disulfide/trisulfide bonding. Circular dichroism spectra assays revealed that H₂S reagents could be likely to react with cysteine thiols to yield sulfurated thiol (-SSH) derivatives in L-LDH, and sulfur-containing PS (polysulfide) was a stronger protein S-sulfurating agent than the other two sulfides. Matrix assisted laser desorptionionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS) study showed partial S-sulfuration of the active cysteine sites existed in L-LDH. In conclusion, H₂S exerts its biological effects as a gasotransmitter through its reactions with cysteine thiols in proteins by S-sulfuration.

Key words:

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1.
沈阳化工大学材料科学与工程学院沈阳 110142

硫化氢信号分子与L-乳酸脱氢酶蛋白相互作用机制的研究

English

Investigation of Interaction Mechanism Between Signaling Molecule Hydrogen Sulfide and L-Lactic Dehydrogenase

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通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

硫化氢信号分子与L-乳酸脱氢酶蛋白相互作用机制的研究

English

Investigation of Interaction Mechanism Between Signaling Molecule Hydrogen Sulfide and L-Lactic Dehydrogenase

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

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