Inactivation of carboxypeptidase Y by mutational removal of the putative essential histidyl residue
- 1 September 1989
- journal article
- research article
- Published by Springer Nature in Carlsberg Research Communications
- Vol. 54 (5) , 165-171
- https://doi.org/10.1007/bf02904470
Abstract
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 · 104 and 7 · 102, 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.This publication has 33 references indexed in Scilit:
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