Characterization of γ-glutamyl kinase mutants from Saccharomyces cerevisiae

Yuki Tatehashi; Hiroshi Takagi

doi:10.1016/j.jbiosc.2013.05.018

Characterization of γ-glutamyl kinase mutants from Saccharomyces cerevisiae

J Biosci Bioeng. 2013 Nov;116(5):576-9. doi: 10.1016/j.jbiosc.2013.05.018. Epub 2013 Jun 14.

Authors

Yuki Tatehashi¹, Hiroshi Takagi

Affiliation

¹ Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.

PMID: 23770122
DOI: 10.1016/j.jbiosc.2013.05.018

Abstract

We characterized the yeast γ-glutamyl kinase important for proline biosynthesis. Asp154 and Ile150 were involved in the affinity with glutamate as well as proline. The Asp154Asn and Ile150Thr mutants had increased thermostability, in addition to desensitization to proline. Deletion of a C-terminal domain dramatically decreased the reaction rate and stability.

Keywords: PUA domain; Proline synthesis; Saccharomyces cerevisiae; Yeast; γ-Glutamyl kinase.

MeSH terms

Amino Acid Sequence
Enzyme Stability / genetics
Glutamic Acid / metabolism
Kinetics
Molecular Sequence Data
Mutant Proteins / genetics
Mutant Proteins / metabolism*
Phosphotransferases (Carboxyl Group Acceptor) / genetics*
Phosphotransferases (Carboxyl Group Acceptor) / metabolism*
Proline / biosynthesis
Proline / metabolism
Protein Structure, Tertiary / genetics
Saccharomyces cerevisiae / enzymology*
Saccharomyces cerevisiae / genetics
Sequence Deletion / genetics

Substances

Mutant Proteins
Glutamic Acid
Proline
Phosphotransferases (Carboxyl Group Acceptor)
glutamate 5-kinase