巯基修饰对中华仓鼠二氢叶酸还原酶的激活.pdfVIP

15 1 Vol. 15, No . 1 1999 2 Chinese Journal of Biochemistry and Molecular Biology Feb. 1999 ·研究简报· 范映辛　李震宇　周筠梅 ( 中国科学院生物物理研究所, 生物大分子国家重点实验室, 北京 100101) Activation of Dihydrofolate Reductase from Chinese Hamster - by Sulfhydryl modification , , FAN Yingxin LI Zhenyu ZHOU Junmei (N ational Laboratory of Biomacromolecules, Institute of Biophysics, Chinese A cademy of Scinence, Beij ing 100101) AbstractThe activity of dihydrofolate reductase from Chinese hamster can be increased 2. 4, 1. 8 and 1. 4 fold upon the modification of its sulfhydryl group w ith p -hydroxylmercuribenzoate (p - ) , - ( - ) 5, 5 - ( 2- ) HM B p hydroxylmercuriphenylsulfonate p HMPS and dithiobis nitrobenzoate acid ( ) . 400- - DT NB respectively T he activation of the enzyme by fold molar excess of p HMB w as a - 1 monophasic process with a rate constant of 0. 015 s . T he extents of activation as w ell as the rate of activation w as low ered in the presence of the substrate dihydrofolate, but w as unaffected by the . other substrate of NADPH The modified enzyme could be further activated by guanidine hy- drochloride and potassium chloride, but not by urea, w hich suggesting that the mechanism of the activation of dihydrofolate reductase by ions w as different from that by protein denaturants and - . sulfhydryl modifyi