The biological role of phosphoproteins depends upon their degree of phosphorylation in vivo. Methods currently available to measure the degree of phosphorylation of a protein involve indirect procedures to detect the 32P-phosphate incorporation. We report here a direct method to measure relative amounts of phospho- and dephospho-forms of peptides based upon a mass spectrometric technique. The intensities of the molecular ions corresponding to the two forms of the peptides are proportional to their relative amounts. This is demonstrated for a peptide fragment of the protein B-50(GAP-43) and for kemptide, respectively substrates for protein kinases C and A, and demonstrates the applicability of fast atom bombardment-mass spectrometry to quantitate peptides bearing post-translational modifications.
Measurement of relative amounts of phospho- and dephospho-B-50(GAP-43) peptides by fast atom bombardment-mass spectrometry / M.M.G. Di Luca, P.N.E. De Graan, L. De Angelis, W.H. Gispen, F. Cattabeni. - In: FEBS LETTERS. - ISSN 0014-5793. - 301:2(1992), pp. 150-154.
Measurement of relative amounts of phospho- and dephospho-B-50(GAP-43) peptides by fast atom bombardment-mass spectrometry
M.M.G. Di LucaPrimo
;F. CattabeniUltimo
1992
Abstract
The biological role of phosphoproteins depends upon their degree of phosphorylation in vivo. Methods currently available to measure the degree of phosphorylation of a protein involve indirect procedures to detect the 32P-phosphate incorporation. We report here a direct method to measure relative amounts of phospho- and dephospho-forms of peptides based upon a mass spectrometric technique. The intensities of the molecular ions corresponding to the two forms of the peptides are proportional to their relative amounts. This is demonstrated for a peptide fragment of the protein B-50(GAP-43) and for kemptide, respectively substrates for protein kinases C and A, and demonstrates the applicability of fast atom bombardment-mass spectrometry to quantitate peptides bearing post-translational modifications.Pubblicazioni consigliate
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