The roles of Glu93 and Tyr149 in astacin‐like zinc peptidases
- 8 November 2000
- journal article
- research article
- Published by Wiley in FEBS Letters
- Vol. 484 (3) , 224-228
- https://doi.org/10.1016/s0014-5793(00)02163-3
Abstract
The catalytic zinc of astacin, a prototype of the astacin family and the metzincin superfamily of metalloproteinases is coordinated by three histidines, a glutamate bound water and a tyrosine. In order to assess the roles of active site key residues, two mutants, Glu93Ala-astacin and Tyr149Phe-astacin, were expressed in Escherichia coli, affinity-purified and renatured. While the Glu93Ala mutant was inactive, the Tyr149Phe mutant retained about 2.5% residual activity toward Dns-Pro-Lys-Arg*Ala-Pro-Trp-Val, based on the k cat/K m value for recombinant wild-type astacin. These results support a model in which Glu93 is the general base in substrate hydrolysis, whereas Tyr149 contributes to transition state binding.Keywords
This publication has 31 references indexed in Scilit:
- Families of zinc metalloproteasesPublished by Wiley ,2001
- Heterologously overexpressed, affinity‐purified human meprin α is functionally active and cleaves components of the basement membrane in vitroFEBS Letters, 1999
- The mechanistic role of the coordinated tyrosine in astacinJournal of Inorganic Biochemistry, 1998
- Site-Directed Mutagenesis of the Active Site Glutamate in Human Matrilysin: Investigation of Its Role in CatalysisBiochemistry, 1997
- The Role of Histidine 231 in Thermolysin-like Enzymes.Journal of Biological Chemistry, 1995
- Astacins, serralysins, snake venom and matrix metalloproteinases exhibit identical zinc‐binding environments (HEXXHXXGXXH and Met‐turn) and topologies and should be grouped into a common family, the ‘metzincins’FEBS Letters, 1993
- Isolation of cDNAs for LCE and HCE, two constituent proteases of the hatching enzyme of Oryzias latipes, and concurrent expression of their mRNAs during developmentDevelopmental Biology, 1992
- Carboxypeptidase AAccounts of Chemical Research, 1989
- Structural basis of the action of thermolysin and related zinc peptidasesAccounts of Chemical Research, 1988