Nuclear magnetic resonance assignments and global fold of a CheY-binding domain in CheA, the chemotaxis-specific kinase of Escherichia coli

Megan McEvoy, Hongjun Zhou, Amy F. Roth, David F. Lowry, Tom B. Morrison, Lewis E. Kay, Frederick W. Dahlquist

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

CheA is the histidine autokinase in the Escherichia coli chemotaxis signal transduction pathway responsible for coupling of signals received by transmembrane receptors to the response regulators CheY and CheB. Here NMR spectroscopy is used to study a 14 kDa fragment of CheA, residues 124-257, that binds the response regulator CheY. Backbone atom resonance assignments were obtained by analysis of 3D HNCACB, 3D CBCA(CO)NH, and HNCO spectra, whereas side-chain assignments were obtained primarily by analysis of 3D H(CCO)NH, 3D C(CO)NH, 3D HCCH-TOCSY, and 3D 1H, 15N TOCSY-HSMQC spectra. NOE cross peak patterns and intensities as well as torsion angle restraints were used to determine the secondary structure, and a low-resolution structure was calculated by hybrid distance-geometry simulated annealing methods. The CheA124-257 fragment consists of four antiparallel β strands and two helices, arranged in an "open-faced β-sandwich" motif, as well as two unstructured ends that correspond to domain linkers in the full-length protein. The 15N-1H correlation spectrum of 15N-labeled CheA124-257 bound to unlabeled CheY shows specific localized changes that may correspond to a CheY-binding face on CheA.

Original languageEnglish (US)
Pages (from-to)13871-13880
Number of pages10
JournalBiochemistry
Volume34
Issue number42
StatePublished - 1995
Externally publishedYes

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Chemotaxis
Carbon Monoxide
Escherichia coli
Phosphotransferases
Magnetic Resonance Spectroscopy
Nuclear magnetic resonance
Signal transduction
Simulated annealing
Histidine
Torsional stress
Nuclear magnetic resonance spectroscopy
Signal Transduction
Atoms
Geometry
Proteins
autophosphorylation-dependent multifunctional protein kinase

ASJC Scopus subject areas

  • Biochemistry

Cite this

McEvoy, M., Zhou, H., Roth, A. F., Lowry, D. F., Morrison, T. B., Kay, L. E., & Dahlquist, F. W. (1995). Nuclear magnetic resonance assignments and global fold of a CheY-binding domain in CheA, the chemotaxis-specific kinase of Escherichia coli. Biochemistry, 34(42), 13871-13880.

Nuclear magnetic resonance assignments and global fold of a CheY-binding domain in CheA, the chemotaxis-specific kinase of Escherichia coli. / McEvoy, Megan; Zhou, Hongjun; Roth, Amy F.; Lowry, David F.; Morrison, Tom B.; Kay, Lewis E.; Dahlquist, Frederick W.

In: Biochemistry, Vol. 34, No. 42, 1995, p. 13871-13880.

Research output: Contribution to journalArticle

McEvoy, M, Zhou, H, Roth, AF, Lowry, DF, Morrison, TB, Kay, LE & Dahlquist, FW 1995, 'Nuclear magnetic resonance assignments and global fold of a CheY-binding domain in CheA, the chemotaxis-specific kinase of Escherichia coli', Biochemistry, vol. 34, no. 42, pp. 13871-13880.
McEvoy, Megan ; Zhou, Hongjun ; Roth, Amy F. ; Lowry, David F. ; Morrison, Tom B. ; Kay, Lewis E. ; Dahlquist, Frederick W. / Nuclear magnetic resonance assignments and global fold of a CheY-binding domain in CheA, the chemotaxis-specific kinase of Escherichia coli. In: Biochemistry. 1995 ; Vol. 34, No. 42. pp. 13871-13880.
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