Phosphorylation causes subtle changes in solvent accessibility at the interdomain interface of methylesterase CheB

Carrie A. Hughes, Jeffrey G. Mandell, Ganesh S. Anand, Ann Stock, Elizabeth A. Komives

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

The crystal structure of the unphosphorylated state of methylesterase CheB shows that the regulatory domain blocks access of substrate to the active site of the catalytic domain. Phosphorylation of CheB at Asp56 results in a catalytically active transiently phosphorylated enzyme with a lifetime of approximately two seconds. Solvent accessibility changes in this transiently phosphorylated state were probed by MALDI-TOF-detected amide hydrogen/deuterium exchange. No changes in solvent accessibility were seen in the regulatory domain upon phosphorylation of Asp56, but two regions in the catalytic domain (199-203 and 310-317) became more solvent accessible. These two regions flank the active site and contain domain-domain contact residues. Comparison with results from the isolated catalytic domain-containing C-terminal fragment of CheB (residues 147-349) showed that the increased solvent accessibility was less than would have occurred upon detachment of the regulatory domain. Thus, phosphorylation causes subtle changes in solvent accessibility at the interdomain interface of CheB.

Original languageEnglish (US)
Pages (from-to)967-976
Number of pages10
JournalJournal of molecular biology
Volume307
Issue number4
DOIs
StatePublished - Apr 6 2001

All Science Journal Classification (ASJC) codes

  • Structural Biology
  • Molecular Biology

Keywords

  • Chemotaxis
  • H/H exchange
  • MALDI-TOF
  • Phosphoaspartate
  • Response regulator

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