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Title: ATP Forms a Stable Complex with the Essential Histidine Kinase WalK (YycG) Domain

Authors:
; ; ; ;
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1158640
Report Number(s):
SLAC-REPRINT-2014-376
DOE Contract Number:
AC02-76SF00515
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallogr. D68: 839-845, 2012
Country of Publication:
United States
Language:
English
Subject:
CHEM

Citation Formats

Celikel, R., Veldore, V.H., Mathews, I., Devine, K.M., and Varughese, K.I.. ATP Forms a Stable Complex with the Essential Histidine Kinase WalK (YycG) Domain. United States: N. p., 2014. Web.
Celikel, R., Veldore, V.H., Mathews, I., Devine, K.M., & Varughese, K.I.. ATP Forms a Stable Complex with the Essential Histidine Kinase WalK (YycG) Domain. United States.
Celikel, R., Veldore, V.H., Mathews, I., Devine, K.M., and Varughese, K.I.. Tue . "ATP Forms a Stable Complex with the Essential Histidine Kinase WalK (YycG) Domain". United States. doi:.
@article{osti_1158640,
title = {ATP Forms a Stable Complex with the Essential Histidine Kinase WalK (YycG) Domain},
author = {Celikel, R. and Veldore, V.H. and Mathews, I. and Devine, K.M. and Varughese, K.I.},
abstractNote = {},
doi = {},
journal = {Acta Crystallogr. D68: 839-845, 2012},
number = ,
volume = ,
place = {United States},
year = {Tue Sep 23 00:00:00 EDT 2014},
month = {Tue Sep 23 00:00:00 EDT 2014}
}
  • The histidine WalK (YycG) plays a crucial role in coordinating murein synthesis with cell division and the crystal structure of its ATP binding domain has been determined. Interestingly the bound ATP was not hydrolyzed during crystallization and remains intact in the crystal lattice. In Bacillus subtilis, the WalRK (YycFG) two-component system coordinates murein synthesis with cell division. It regulates the expression of autolysins that function in cell-wall remodeling and of proteins that modulate autolysin activity. The transcription factor WalR is activated upon phosphorylation by the histidine kinase WalK, a multi-domain homodimer. It autophosphorylates one of its histidine residues by transferringmore » the γ-phosphate from ATP bound to its ATP-binding domain. Here, the high-resolution crystal structure of the ATP-binding domain of WalK in complex with ATP is presented at 1.61 Å resolution. The bound ATP remains intact in the crystal lattice. It appears that the strong binding interactions and the nature of the binding pocket contribute to its stability. The triphosphate moiety of ATP wraps around an Mg{sup 2+} ion, providing three O atoms for coordination in a near-ideal octahedral geometry. The ATP molecule also makes strong interactions with the protein. In addition, there is a short contact between the exocyclic O3′ of the sugar ring and O2B of the β-phosphate, implying an internal hydrogen bond. The stability of the WalK–ATP complex in the crystal lattice suggests that such a complex may exist in vivo poised for initiation of signal transmission. This feature may therefore be part of the sensing mechanism by which the WalRK two-component system is so rapidly activated when cells encounter conditions conducive for growth.« less
  • The 2 equiv of divalent cation that are required cofactors for pyruvate kinase reside in sites of different affinities for different species of cation. The intrinsic selectivity of the protein-based site for Mn(II) and of the nucleotide-based site for Mg(II) has been exploited in electron paramagnetic resonance (EOR) investigations of ligands for Mn(II) at the protein-based site. Oxalate, a structural analogue of the enolate of pyruvate, has been used as a surrogate for the reactive form of pyruvate in complexes with enzyme, Mn(II), Mg(II), and ATP. Superhyperfine coupling between the unpaired electron spin of Mn(II) and the nuclear spin ofmore » /sup 17/O, specifically incorporated into oxalate, shows that oxalate is bound at the active site as a bidentate chelate with Mn(II). Coordination of the ..gamma..-phosphate of ATP to this same Mn(II) center is revealed by observation of superhyperfine coupling from /sup 17/O regiospecifically incorporated into the ..gamma..-phosphate group of ATP. By contrast, /sup 17/O in the ..cap alpha..-phosphate or in the ..beta..-phosphate groups of ATP does not influence the spectrum. Experiments in /sup 17/O-enriched water show that there is also a single water ligand bound to the Mn(II). These data indicate that ATP bridges Mn(II) and Mg(II) at the active site. A close spacing of the two divalent cations is also evident from the occurrence of magnetic interactions for complexes in which 2 equiv of Mn(II) are present at the active site. The structure for the enzyme-Mn(II)-oxalate-Mg(II)-ATP complex suggests a scheme for the normal reverse reaction of pyruvate kinase in which the divalent cation at the protein-based site activates the keto acid substrate through chelation and promotes phospho transfer by simultaneous coordination to the enolate oxygen and to a pendant oxygen from the ..gamma..-phosphate of ATP.« less
  • No abstract prepared.
  • Histidine kinase receptors are elements of the two-component signal transduction systems commonly found in bacteria and lower eukaryotes, where they are crucial for environmental adaption through the coupling of extracellular changes to intracellular responses. The typical two-component system consists of a membrane-spanning histidine kinase sensor and a cytoplasmic response regulator. In the calssic system, extracellular signals such as small molecule ligands and ions are detected by the periplasmic sensor domain of the histidine kinase receptor, which modulates the catalytic activity of the cytoplasmic histidine kinase domain and promotes ATP-dependent autophosphorylation of a conserved histidine residue. G. sulfurreducens genomic DNA wasmore » used.« less