溴化十烷基三甲铵对肌酸激酶的变性及复性

测定了溴化十烷基三甲铵(C_(10)H_(21)N(CH_3)_3Br;记为C_(10)NM_3)对肌酸激酶(Creatine Kinase;记为C.K.)的活力及构象的影响,以及变性后C.K.的复性。实验结果表明:C_(10)NM_3对C.K.有很强的变性能力,在0.06M时,就可以使C.K.完全失活;在0.08M时,就可使C.K.内部的6个巯基有4个暴露出来;与SDS变性剂不同,C.K.在高浓度的C_(10)NM_3中变性以后,直接冲稀时就可以完全复性。FTIR以及CD等实验方法证明,尽管C_(10)NM_3能使C.K.的构象发生明显变化,但C.K.的二级结构几乎不受C_(10)NM_3的影响。

THE DENATURATION AND RENATURATION OF CREATINE KINASE BY DECYLTRIMETHYL AMMONIUM BROMIDE

The effect of decyltri methyl ammonium bromide C10H21N(CH3)3Br (C10NM3) on the activity of creatine kinase (C.K.), and the renaturation of denatured C.K have been studied. The results show that the denaturation ability of C10 NM3 is very high.When the concentration of C10NM3 is 0.06M, C.K. loses its enzyme activity entirely. When the concentration of C10NM3 is 0.08M, 4 out of 6 thiol buried in inner of C.K. can be measured. When C.K. is denatured at a high concentration of C10NM3, its activity can be entirely recovered after dilution the solution directly, which is different from that of SDS. The results of FTIR and CD show that the secondary structure of C.K. has not been affected although the conformation of C.K. has been changed very much when the concentration of C10NM3 is higher than 0.07M.