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The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS

The Escherichia coli CheY protein belongs to a large bacterial response regulator superfamily. X-ray hydroxy radical foot-printing with mass spectroscopy (XFMS) has shown that allosteric activation of CheY by its motor target triggers a concerted internalization of aromatic sidechains. We reanalyzed...

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Autores principales: Hamid, Maham, Khalid, Muhammad Farhan, Chaudhary, Safee Ullah, Khan, Shahid
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9659070/
https://www.ncbi.nlm.nih.gov/pubmed/36361564
http://dx.doi.org/10.3390/ijms232112771
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author Hamid, Maham
Khalid, Muhammad Farhan
Chaudhary, Safee Ullah
Khan, Shahid
author_facet Hamid, Maham
Khalid, Muhammad Farhan
Chaudhary, Safee Ullah
Khan, Shahid
author_sort Hamid, Maham
collection PubMed
description The Escherichia coli CheY protein belongs to a large bacterial response regulator superfamily. X-ray hydroxy radical foot-printing with mass spectroscopy (XFMS) has shown that allosteric activation of CheY by its motor target triggers a concerted internalization of aromatic sidechains. We reanalyzed the XFMS data to compare polar versus non-polar CheY residue positions. The polar residues around and including the 57D phosphorylated site had an elevated hydroxy radical reactivity. Bioinformatic measures revealed that a water-mediated hydrogen bond network connected this ring of residues with the central 57D. These residues solvated 57D to energetically stabilize the apo-CheY fold. The abundance of these reactive residues was reduced upon activation. This result was supported by the bioinformatics and consistent with the previously reported activation-induced increase in core hydrophobicity. It further illustrated XFMS detection of structural waters. Direct contacts between the ring residues and the phosphorylation site would stabilize the aspartyl phosphate. In addition, we report that the ring residue, 18R, is a constant central node in the 57D solvation network and that 18R non-polar substitutions determine CheY diversity as assessed by its evolutionary trace in bacteria with well-studied chemotaxis. These results showcase the importance of structured water dynamics for phosphorylation-mediated signal transduction.
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spelling pubmed-96590702022-11-15 The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS Hamid, Maham Khalid, Muhammad Farhan Chaudhary, Safee Ullah Khan, Shahid Int J Mol Sci Article The Escherichia coli CheY protein belongs to a large bacterial response regulator superfamily. X-ray hydroxy radical foot-printing with mass spectroscopy (XFMS) has shown that allosteric activation of CheY by its motor target triggers a concerted internalization of aromatic sidechains. We reanalyzed the XFMS data to compare polar versus non-polar CheY residue positions. The polar residues around and including the 57D phosphorylated site had an elevated hydroxy radical reactivity. Bioinformatic measures revealed that a water-mediated hydrogen bond network connected this ring of residues with the central 57D. These residues solvated 57D to energetically stabilize the apo-CheY fold. The abundance of these reactive residues was reduced upon activation. This result was supported by the bioinformatics and consistent with the previously reported activation-induced increase in core hydrophobicity. It further illustrated XFMS detection of structural waters. Direct contacts between the ring residues and the phosphorylation site would stabilize the aspartyl phosphate. In addition, we report that the ring residue, 18R, is a constant central node in the 57D solvation network and that 18R non-polar substitutions determine CheY diversity as assessed by its evolutionary trace in bacteria with well-studied chemotaxis. These results showcase the importance of structured water dynamics for phosphorylation-mediated signal transduction. MDPI 2022-10-23 /pmc/articles/PMC9659070/ /pubmed/36361564 http://dx.doi.org/10.3390/ijms232112771 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hamid, Maham
Khalid, Muhammad Farhan
Chaudhary, Safee Ullah
Khan, Shahid
The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS
title The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS
title_full The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS
title_fullStr The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS
title_full_unstemmed The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS
title_short The Solvation of the E. coli CheY Phosphorylation Site Mapped by XFMS
title_sort solvation of the e. coli chey phosphorylation site mapped by xfms
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9659070/
https://www.ncbi.nlm.nih.gov/pubmed/36361564
http://dx.doi.org/10.3390/ijms232112771
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