对生蒴蛤谷胱甘肽转移酶的光谱研究

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.

Spectroscopic Characterization of Glutathione S-Transferase of Asaphis dichotoma

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.