Determination of Phosphohistidine Stoichiometry in Histidine Kinases by Intact Mass Spectrometry

Tomlinson, Lauren J, Coldron, Alice KM Clubbs, Eyers, Patrick A ORCID: 0000-0002-9220-2966 and Eyers, Claire E ORCID: 0000-0002-3223-5926 (2020) Determination of Phosphohistidine Stoichiometry in Histidine Kinases by Intact Mass Spectrometry. HISTIDINE PHOSPHORYLATION, 2077. pp. 83-91.

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Official URL: http://dx.doi.org/10.1007/978-1-4939-9884-5_6

Abstract

Protein histidine phosphorylation has largely remained unexplored due to the challenges of analyzing relatively unstable phosphohistidine-containing proteins. We describe a procedure for determining the stoichiometry of histidine phosphorylation on the human histidine kinases NME1 and NME2 by intact mass spectrometry under conditions that retain this acid-labile protein modification. By characterizing these two model histidine protein kinases in the absence and presence of a suitable phosphate donor, the stoichiometry of histidine phosphorylation can be determined. The described method can be readily adapted for the analysis of other proteins containing phosphohistidine.

Item Type:	Article
Uncontrolled Keywords:	NME1, NME2, Histidine phosphorylation, Intact mass spectrometry
Depositing User:	Symplectic Admin
Date Deposited:	02 Dec 2019 09:35
Last Modified:	19 Jan 2023 00:17
DOI:	10.1007/978-1-4939-9884-5_6
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3064396