Study of Secondary Specificity of Enteropeptidase in Comparison with Trypsin
Study of Secondary Specificity of Enteropeptidase in Comparison with Trypsin
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肠肽酶与胰蛋白酶的二级特异性比较研究
DOI：
10.1023/b:biry.0000040224.47278.3b
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
L. Rumsh
中科院分区：
文献类型：
--
作者：
A. G. Mikhailova;V. V. Likhareva;B. V. Vaskovsky;S. K. Garanin;L. V. Onoprienko;I. Prudchenko;L. Chikin;L. Rumsh
使用通式为A-（Asp/Glu）n-Lys（Arg）-↓-B（n = 1-4）的肽底物，对肠肽酶和胰蛋白酶的二级特异性进行了比较研究。这是第一个研究证明，类似于其他丝氨酸蛋白酶，肠肽酶具有延伸的二级结合位点，与待水解的肽键周围的6-7个氨基酸残基相互作用。然而，在P2-P5位含有四个带负电荷的Asp/Glu残基的典型肠肽酶底物的情况下，这些残基与肠肽酶轻链的二级位点Lys 99之间的静电相互作用是决定水解效率的主要因素。肠肽酶的第二特异性不同于胰蛋白酶的第二特异性。产酶底物G5 DK-F（NO2）G（kcat/Km = 2380 mM-1·min-1）比融合蛋白PrAD 4K-P26（kcat/Km = 1260 mM-1·min-1）更有效。
A comparative study of secondary specificities of enteropeptidase and trypsin was performed using peptide substrates with general formula A-(Asp/Glu)n-Lys(Arg)-↓-B, where n = 1-4. This was the first study to demonstrate that, similar to other serine proteases, enteropeptidase has an extended secondary binding site interacting with 6-7 amino acid residues surrounding the peptide bond to be hydrolyzed. However, in the case of typical enteropeptidase substrates containing four negatively charged Asp/Glu residues at positions P2-P5, electrostatic interaction between these residues and the secondary site Lys99 of the enteropeptidase light chain is the main factor that determines hydrolysis efficiency. The secondary specificity of enteropeptidase differs from the secondary specificity of trypsin. The chromophoric synthetic enteropeptidase substrate G5DK-F(NO2)G (kcat/Km = 2380 mM–1·min–1) is more efficient than the fusion protein PrAD4K-P26 (kcat/Km = 1260 mM–1·min–1).
影响因子：
5.6
作者：
Lu, DS;Fütterer, K;Sadler, JE
通讯作者：
Sadler, JE
影响因子：
6.5
作者：
W. Safi;J. N. Maiorano;W. Davidson
通讯作者：
W. Safi;J. N. Maiorano;W. Davidson
影响因子：
2.9
作者：
Volker Schellenberger;Christoph W. Turck;William J. Rutter
通讯作者：
Volker Schellenberger;Christoph W. Turck;William J. Rutter