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Anal Bioanal Chem. 2010 Aug;397(8):3209-12. doi: 10.1007/s00216-009-3372-x. Epub 2010 Jan 10.

Stepchild phosphohistidine: acid-labile phosphorylation becomes accessible by functional proteomics.

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Integrierte Funktionelle Genomik, Interdisziplinäres Zentrum für Klinische Forschung, Westfälische Wilhelms-Universität Münster, Roentgenstrasse 21, 48149 Münster, Germany.


Bioanalytical techniques were preferentially developed for the investigation of phosphohydroxyamino acids in the past and there is a wealth of information on the detection of serine, threonine and tyrosine phosphorylation in functional proteomics. However, similarly important for protein regulation and signalling is the phosphorylation of other amino acids such as histidine, but its detection is hampered by the sensitivity to acid. Mass spectrometry in conjunction with chromatographic methods is allowing us to start to get a handle on phosphohistidine. (32)P-labelling and amino acid analysis for phosphorylation site determination is increasingly complemented by typical proteomic approaches based on reversed-phase peptide separation and gas-phase fragmentation. Chemical phosphorylation of peptides is a valuable tool, therefore, for the generation of analytical standards for use in method development.

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