对生蒴蛤谷胱甘肽转移酶的光谱研究

高等学校化学学报 ›› 2003, Vol. 24 ›› Issue (8): 1472.

对生蒴蛤谷胱甘肽转移酶的光谱研究

杨海灵, 高波, 曾庆银, 付学奇, 聂力嘉, 朱圣庚, 周先碗

1.北京大学生命科学学院, 北京 100871;
2. 吉林大学生命科学学院, 长春 130023

收稿日期:2002-04-28 出版日期:2003-08-24 发布日期:2003-08-24
通讯作者: 周先碗(1949年出生),男,教授,博士生导师,从事蛋白质结构与功能研究.E-mail:xwzhou@pku.edu.cn付学奇(1960年出生),男,博士,教授,从事细胞信号传导与药物筛选研究.E-mail:fxq@mail.jlu.edu.cn E-mail:xwzhou@pku.edu.cn; fxq@mail.jlu.edu.cn
基金资助:
国家海洋“八六三”计划(批准号:819-04-02)资助

Spectroscopic Characterization of Glutathione S-Transferase of Asaphis dichotoma

YANG Hai-Ling, GAO Bo, ZENG Qing-Yin, FU Xue-Qi, NIE Li-Jia, ZHU Sheng-Geng, ZHOU Xian-Wan

1. College of Life Sciences, Peking University, Beijing, 100871, China;
2. College of Life Sciences, Jilin University, Changchun 130023, China

Received:2002-04-28 Online:2003-08-24 Published:2003-08-24

摘要/Abstract

摘要： 为深入了解海洋软体生物谷胱甘肽转移酶(GSTs)的性质和特点,以对生蒴蛤肝肠为材料,经GSH亲和层析与反向HPLC分离得到纯化蛋白(adGST),经SDS-PAGE、凝胶过滤与飞行时间质谱分析确定该酶为双亚基组成,全酶分子量50000,亚基为25000.紫外光谱分析表明,该酶在280nm有最大吸收峰;荧光光谱分析表明,该酶的最大激发波长为280nm,在350nm有最大发射峰;用圆二色谱与傅里叶变换红外光谱对该酶进行二级结构测定,结果表明,该酶含有约35%左右的α-螺旋,30%左右的β-折叠,表明该酶属于典型的球状蛋白.

关键词: 谷胱甘肽转移酶(GSTs), 圆二色谱, 傅里叶变换红外光谱, 二级结构

Abstract: AnoveLIsoenzyme of glutathione S-transferase(adGST) was purified from the liver intestine of the seashell(Asaphis dichotoma) by GST-Sepharose 4 Baffinity chromatography and reverse HPLC.The enzyme was composed of two subunits with a molecular weight of 23000 for each determined by SDS-PAGE, MALDITOF-MSand gel filtration.The characteristic peak of ultraviolet absorbance spectra of the ad GSTwas at 280 nm, and that of fluorescence excitation spectrum was at 280 nm and that of the fluorescence emission spectrum was at 350 nm.The CDspectra and FTIRspectroscopy show that the ad GSThas a total secondary structure content of about 35% α-helix and 30% sheets.The adGSTwas a kind of global protein.

Key words: Glutathione S-transferase(adGST), CD spectrum, FTIR spectrum, Secondary structure

中图分类号:

TrendMD:

杨海灵, 高波, 曾庆银, 付学奇, 聂力嘉, 朱圣庚, 周先碗. 对生蒴蛤谷胱甘肽转移酶的光谱研究. 高等学校化学学报, 2003, 24(8): 1472.

YANG Hai-Ling, GAO Bo, ZENG Qing-Yin, FU Xue-Qi, NIE Li-Jia, ZHU Sheng-Geng, ZHOU Xian-Wan. Spectroscopic Characterization of Glutathione S-Transferase of Asaphis dichotoma. Chem. J. Chinese Universities, 2003, 24(8): 1472.

参考文献

[1] Armstrong R.N..Chem.Res.Toxicol.[J],1991,4(2):131—140 [2] Cheng Yuan-Kai(程元恺).Prog.Biochem.Biophys.(生物化学与生物物理进展)[J],1994,21(5):395—399 [3] Morgan A.S.,Ciaccio P.J.,Tew K.D..Cancer Chemotherapyand Pharmacology[J],1996,37(4):363—370 [4] Hodge S.,Longley M.,Booth L.etal..Bull.Environ.Contam.Toxicol.[J],2000,65:8—15 [5] Sato Y.,Boger P.,Wadabayashi K..J.Pestic.Sci.[J],1996,22(1):33—36 [6] LI Chun-Hai(李春海),T A N Zi-Xin(谭子兴).Chinese Biochemical Journal(生物化学杂志)[J],1989,5(1):78—84 [7] M A Su-Yong(马素永),ZHO U Xian-Wan(周先碗),ZHANG Ting-Fang(张庭芳).Acta Scientiarum Naturalium UniversitatisPekinensis(北京大学学报,自然科学版)[J],1999,35(4):474—479 [8] ZHO U Xian-Wan(周先碗).Chinese Journalof Biochemistry and Molecular Biology(生物化学与分子生物学报)[J],1999,15(2):269—273 [9] ZHANG Feng(张锋),SO N G Xi-Ming(宋溪明),LIOU Qi-Tao(刘祁涛).Chem.J.Chinese Universities(高等学校化学学报)[J],2000,21(9):1 348—1 352 [10] LUO Shi-Zhong(罗施中),ZHA O Gang(赵刚),LI Yan-Mei(李艳梅)etal..Chem.J.Chinese Universities(高等学校化学学报)[J],2002,23(5):852—854—1 133 [11] SU N Yu-Long(孙雨龙),WANG Yun-Hua(王韵华),ZHO U Gang(周刚)et al..Chem.J.Chinese Universities(高等学校化学学报)[J],1996,17(11):1 673—1 677 [12] WANG Yong-Ting(王永亭),HUANG Pin-Wei(黄品伟),Li Zheng-Qing(李正强)etal..Chem.J.Chinese Universities(高等学校化学学报)[J],2001,22(7):1 117—1 119 [13] JIA N G Jun-Guang(蒋俊光),WANG Zhen-Xin(王振新),LIU Chang-Wei(刘长伟)etal..Chem.J.Chinese Universities(高等学校化学学报)[J],2001,22(7):1 131—1 133 [14] Mannervik B.,Guthenberg C.;Ed.Jakoby W.B..Glutathione Transferase(Human Placenta),Methods Enzymology[M],New York:Academic Press,1981,77:231—235 [15] Habig W.H.,Pabst M.J.,Jakoby W.B..J.Biol.Chem.[J],1974,249(22):7 130—7 139 [16] Lowry O.H..J.Biol.Chem.[J],1951,193:265—275 [17] ZHANG Long-Xiang(张龙翔),ZHA G N Ting-Fang(张庭芳),LI Ling-Yuan(李令媛).Experimental Methodsand TechniquesinBiochemistry(生化实验方法和技术)[M],Beijing:Advanced Education Press,1997:122—140 [18] S HEN Tong(沈同),WANG Qing-Yan(王镜岩).Biochemist ry(生物化学)[M],Beijing:Advanced Education Press,1990:85—88 [19] Thomas P.A.,Walter J.,Wolf Y.etal..Applied Spectroscopy[J],1991,45:1 665—1 671 [20] Jagdeesh Bandekar.Biochim.Biophys.Acta[J],1992,1120:123—143