A monomeric histidine kinase derived from EnvZ, an Echerichia coli osmosensor

Ling Qin, Rinku Dutta, Hirofumi Kurokawa, Mitsuhiko Ikura, Masayori Inouye

研究成果: Article査読

31 被引用数 (Scopus)

抄録

Histidine kinases function as dimers. The kinase domain of the osmosensing histidine kinase EnvZ of Escherichia coli consists of two domains: domain A (67 residues) responsible for histidine phosphotransfer and dimerization, and domain B (161 residues) responsible for the catalytic and ATP-binding function. The individual structures of these two domains have been recently solved by NMR spectroscopy. Here, we demonstrate that an enzymatically functional monomeric histidine kinase can be constructed by fusing in tandem two domains A and one domain B to produce a single polypeptide (A-A-B). We show that this protein, EnvZc[AAB], is soluble and exists as a stable monomer. The autophosphorylation and OmpR kinase activities of the monomeric EnvZc[AAB] are similar to that of the wild-type EnvZ, while OmpR-binding and phosphatase functions are reduced. V8 protease digestion and mutational analyses indicate that His-243 of only the amino proximal domain A is phosphorylated. Based on these results, molecular models are proposed for the structures of EnvZc[AAB] and the kinase domain of EnvZ. The present results demonstrate for the first time the construction of a functional, monomeric histidine kinase, further structural studies of which may provide important insights into the structure-function relationships of histidine kinases.

本文言語English
ページ(範囲)24-32
ページ数9
ジャーナルMolecular Microbiology
36
1
DOI
出版ステータスPublished - 2000
外部発表はい

ASJC Scopus subject areas

  • 微生物学
  • 分子生物学

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