Protein Phosphorylation Degree:  Determination by Capillary Liquid Chromatography and Inductively Coupled Plasma Mass Spectrometry

Capillary liquid chromatography (μLC) interfaced to inductively coupled plasma mass spectrometry (ICPMS) is introduced as a new micromethod to determine the phosphorylation degree in phosphoproteins and phosphopeptides containing cysteine and/or methionine residues. The stoichiometric phosphorus to...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2001-07, Vol.73 (13), p.3006-3010
Main Authors: Wind, Mathias, Wesch, Horst, Lehmann, Wolf D
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Catalytic Domain
Chromatography
Chromatography, Liquid - methods
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Mass Spectrometry - methods
Molecular Sequence Data
Phosphoproteins - chemistry
Phosphoproteins - metabolism
Phosphorylation
Proteins
Spectrum analysis
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Summary:Capillary liquid chromatography (μLC) interfaced to inductively coupled plasma mass spectrometry (ICPMS) is introduced as a new micromethod to determine the phosphorylation degree in phosphoproteins and phosphopeptides containing cysteine and/or methionine residues. The stoichiometric phosphorus to sulfur (31P to 32S) ratio is experimentally determined by μLC−ICPMS and converted into the degree of phosphorylation using protein/peptide sequence information. The method is applied to the phosphoproteins α-casein, β-casein, and recombinant protein kinase A catalytic subunit and to synthetic phosphopeptides. The accurate data obtained by μLC−ICPMS allow quantitative assessment of the compound-specific discrimination of the electrospray ionization process between nonphosphorylated and phosphorylated proteins and peptides.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac010066s