Mechanism of CheA protein kinase activation in receptor signaling complexes

Mikhail N. Levit, Yi Liu, Jeffry B. Stock

Research output: Contribution to journalArticlepeer-review

47 Scopus citations


The chemotaxis receptor for aspartate, Tar, generates responses by regulating the activity of an associated histidine kinase, CheA. Tar is composed of an extracellular sensory domain connected by a transmembrane sequence to a cytoplasmic signaling domain. The cytoplasmic domain fused to a leucine zipper dimerization domain forms soluble active ternary complexes with CheA and an adapter protein, CheW. The kinetics of kinase activity within these complexes compared to CheA alone indicate approximately a 50% decrease in the K(M) for ATP and a 100-fold increase in the V(max). A truncated CheA construct that lacks the phosphoaccepting H-domain and the CheY/CheB-binding domain forms an activated ternary complex that is similar to the one formed by the full-length CheA protein. The V(max) of H-domain phosphorylation by this complex is enhanced approximately 60-fold, the K(M) for ATP decreased to 50%, and the K(M) for H-domain decreased to 20% of the values obtained with the same CheA construct in the absence of receptor and CheW. The kinetic data support a mechanism of CheA regulation that involves perturbation of an equilibrium between an inactive form where the H-domain is loosely bound and an active form where the H-domain is tightly associated with the CheA active site and properly positioned for phosphotransfer. The data are consistent with an asymmetric mechanism of CheA activation [Levit, M., Liu, I., Surette, M. G., and Stock, J. B. (1996) J. Biol. Chem. 271, 32057-32063] wherein only one phosphoaccepting domain of CheA at a time can interact with an active center within a CheA dimer.

Original languageEnglish (US)
Pages (from-to)6651-6658
Number of pages8
Issue number20
StatePublished - May 18 1999

All Science Journal Classification (ASJC) codes

  • Biochemistry


Dive into the research topics of 'Mechanism of CheA protein kinase activation in receptor signaling complexes'. Together they form a unique fingerprint.

Cite this