通过突变去除假定的必需组氨酸残基而使羧基肽酶Y失活。

L M Bech, K Breddam
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引用次数: 24

摘要

羧基肽酶Y是一种丝氨酸羧基肽酶,被认为含有类似于丝氨酸内肽酶的催化三联体。根据各种丝氨酸羧基肽酶之间的同源性,推测His-397是催化三联体的一部分。为了验证这一点,将克隆的PRC1基因通过定点诱变与Ala和Arg交换。突变酶的催化效率分别降低了2 × 10(4)和7 × 10(2),证实了His-397在催化中的关键作用。用Hg++或CNBr处理α -397- cpd - y,从而修饰位于活性位点附近的Cys-341,消除了酶的残留活性,表明该残基参与了催化作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Inactivation of carboxypeptidase Y by mutational removal of the putative essential histidyl residue.

Carboxypeptidase Y is a serine carboxypeptidase assumed to contain a catalytic triad similar to the serine endopeptidases. On the basis of the homology between various serine carboxypeptidases His-397 is suspected to be part of the catalytic triad. To test this it was exchanged with Ala and Arg by site-directed mutagenesis of the cloned PRC1 gene. The catalytic efficiency of the mutant enzymes were reduced by a factor of 2 X 10(4) and 7 X 10(2), respectively, confirming the key role of His-397 in catalysis. Treatment of Ala-397-CPD-Y with Hg++ or CNBr, hence modifying Cys-341 located in the vicinity of the active site abolished the residual activity of the enzyme, indicating an additional involvement of this residue in catalysis.

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