一种新的具有纤溶活性的N-V蛋白酶酶学性质分析

doi:吉林省科技厅科技发展计划项目资助课题（20

一种新的具有纤溶活性的N-V蛋白酶酶学性质分析

李奇¹，王昭²，张纪周¹，洪敏¹

1．吉林大学基础医学院生物化学教研室，吉林长春 130021；2.佳木斯大学基础医学院生物化学教研室,黑龙江佳木斯 154000

收稿日期:2007-10-10 修回日期:1900-01-01 出版日期:2008-05-28 发布日期:2008-05-28
通讯作者: 洪敏

Analysis of enzymological characterization of N-V proteinase with fibrinolytic activity

LI Qi¹, WANG Zhao², ZHANG Ji-zhou¹，HONG Min¹

1.Department of Biochemistry, School of Basic Medical Sciences, Jilin University, Changchun 130021 , China;2.Department of Biochemistry, School of Basic Medical Sciences, Jiamusi University， Jiamusi 154000,China

Received:2007-10-10 Revised:1900-01-01 Online:2008-05-28 Published:2008-05-28
Contact: HONG Min

摘要/Abstract

摘要： 目的：测定N-V蛋白酶的最适反应温度、最适反应pH值及酶促反应动力学常数（Km），并对其进行蛋白酶分类。方法：利用纤维蛋白平板法测定N-V蛋白酶最适反应温度、最适反应pH值。以L-亮氨酸-p-硝基苯胺酯(Leu-pNA)作为底物，测定N-V蛋白酶的Km值。利用L-反式环氧琥珀酰亮氨酰氨基（4-胍基）丁烷（E64）、苯甲基磺酰氟（PMSF）、1,10-邻菲啰啉、抑肽酶作为抑制剂对N-V蛋白酶的纤溶活性进行抑制作用，确定其蛋白酶分类。结果：N-V蛋白酶最适反应温度为50℃，最适反应pH值为8～9, Km值为1.97×10^-3mol •L^-1 ，PMSF有效抑制N-V蛋白酶的纤溶活性(P<0.01），而E64、1,10-邻菲啰啉啉、抑肽酶对其活性均无明显抑制作用(P>0.05）。结论： N-V蛋白酶属于丝氨酸蛋白酶类。

关键词: Km值, 丝氨酸内肽酶类

Abstract: Abstract:Objective To detect the optimum reaction temperature ,pH, Km of N-V proteinase and determine its enzymological classification.Methods The fibrin plate method was used to determine the optimum reaction temperature and pH of N-V proteinase.The Km of N-V proteinase was detected by using Leu-pNA as substrate.E64, PMSF, zymofren and 1,10-phenanthroline were used as inhibitors to determine proteinase classification of N-V proteinase.Results The optimum reaction temperature was 50℃, pH was 8-9,Km was 1.9710^-3mol•L^-1.PSMF inhibited the fibrinolytic activity of N-V proteinase （P<0.01）, whereas E64, zymofren and 1,10-phenanthroline had not significantly inhibitory effects on its activity（P>0.05）.Conclusion N-V proteinase belongs to serine proteinase.

Key words: Km, serine endopetidases

中图分类号:

Q556.3

李奇，王昭，张纪周，洪敏. 一种新的具有纤溶活性的N-V蛋白酶酶学性质分析[J]. J4, 2008, 34(3): 377-380.

LI Qi, WANG Zhao, ZHANG Ji-zhou，HONG Min. Analysis of enzymological characterization of N-V proteinase with fibrinolytic activity[J]. J4, 2008, 34(3): 377-380.