Probing Catalytically Essential Domain Orientation in Histidine Kinase EnvZ by Targeted Disulfide Crosslinking

Probing Catalytically Essential Domain Orientation in Histidine Kinase EnvZ by Targeted Disulfide... EnvZ, a dimeric transmembrane histidine kinase, belongs to the family of His-Asp phosphorelay signal transduction systems. The cytoplasmic kinase domain of EnvZ can be dissected into two independently functioning domains, A and B, whose NMR solution structures have been individually determined. Here, we examined the topological arrangement of these two domains in the EnvZ dimer, a structure that is key to understanding the mechanism underlying the autophosphorylation activity of the kinase. A series of cysteine substitution mutants were constructed to test the feasibility of chemical crosslinking between the two domains. These crosslinking data demonstrate that helix I of domain A of one subunit in the EnvZc dimer is in close proximity to domain B of the other subunit in the same dimer, while helix II of domain A of one subunit interacts with domain B of the same subunit in the EnvZc dimer. This is the first demonstration of the topological arrangement between the central dimerization domain containing the active center His residues (domain A) and the ATP-binding catalysis assisting domain (domain B) in a class I histidine kinase. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Molecular Biology Elsevier

Probing Catalytically Essential Domain Orientation in Histidine Kinase EnvZ by Targeted Disulfide Crosslinking

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Publisher
Elsevier
Copyright
Copyright © 2003 Elsevier Science Ltd
ISSN
0022-2836
DOI
10.1016/S0022-2836(03)00275-4
Publisher site
See Article on Publisher Site

Abstract

EnvZ, a dimeric transmembrane histidine kinase, belongs to the family of His-Asp phosphorelay signal transduction systems. The cytoplasmic kinase domain of EnvZ can be dissected into two independently functioning domains, A and B, whose NMR solution structures have been individually determined. Here, we examined the topological arrangement of these two domains in the EnvZ dimer, a structure that is key to understanding the mechanism underlying the autophosphorylation activity of the kinase. A series of cysteine substitution mutants were constructed to test the feasibility of chemical crosslinking between the two domains. These crosslinking data demonstrate that helix I of domain A of one subunit in the EnvZc dimer is in close proximity to domain B of the other subunit in the same dimer, while helix II of domain A of one subunit interacts with domain B of the same subunit in the EnvZc dimer. This is the first demonstration of the topological arrangement between the central dimerization domain containing the active center His residues (domain A) and the ATP-binding catalysis assisting domain (domain B) in a class I histidine kinase.

Journal

Journal of Molecular BiologyElsevier

Published: Apr 25, 2003

References

  • Signal transduction via the histidyl-aspartyl phosphorelay
    Egger, L.A.; Park, H.; Inouye, M.
  • A monomeric histidine kinase derived from EnvZ, an Escherichia coli osmosensor
    Qin, L.; Dutta, R.; Kurokawa, H.; Ikura, M.; Inouye, M.
  • Communication modules in bacterial signaling proteins
    Parkinson, J.S.; Kofoid, E.C.
  • Cysteine scanning mutagenesis of putative transmembrane helices IX and X in the lactose permease of Escherichia coli
    Sahin-Toth, M.; Kaback, H.R.

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