Divalent Metal Ion Binding to the CheY Protein and Its Significance to Phosphotransfer in Bacterial Chemotaxis

Gudrun S. Lukat, Ann M. Stock, Jeffry B. Stock

Research output: Contribution to journalArticlepeer-review

169 Scopus citations

Abstract

Signal transduction in bacterial Chemotaxis involves transfer of a phosphoryl group between the cytoplasmic proteins CheA and CheY. In addition to the established metal ion requirement for autophosphorylation of CheA, divalent magnesium ions are necessary for the transfer of phosphate from CheA to CheY. The work described here demonstrates via fluorescence studies that CheY contains a magnesium ion binding site. This site is a strong candidate for the metal ion site required to facilitate phosphotransfer from phospho-CheA to CheY. The diminished magnesium ion interaction with CheY mutant D13N and the lack of metal ion binding to D57N along with significant reduction in phosphotransfer to these two mutants are in direct contrast to the behavior of wild-type CheY. This supports the hypothesis that the acidic pocket formed by Aspl3 and Asp57 is essential to metal binding and phosphotransfer activity. Metal ion is also required for the dephosphorylation reaction, raising the possibility that the phosphotransfer and hydrolysis reactions occur by a common metal-phosphoprotein transition-state intermediate. The highly conserved nature of the proposed metal ion binding site and site of phosphorylation within the large family of phosphorylated regulatory proteins that are homologous to CheY supports the hypothesis that all these proteins function by a similar catalytic mechanism.

Original languageEnglish (US)
Pages (from-to)5436-5442
Number of pages7
JournalBiochemistry
Volume29
Issue number23
DOIs
StatePublished - Jun 1 1990

All Science Journal Classification (ASJC) codes

  • Biochemistry

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