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The use of mass spectrometry to examine the formation and hydrolysis of the phosphorylated form of phosphoglycerate mutase
Author(s) -
Nairn Jacqueline,
Krell Tino,
Coggins John R.,
Pitt Andrew R.,
Fothergill-Gilmore Linda A.,
Walter Rebecca,
Price Nicholas C.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)00044-a
Subject(s) - phosphoglycerate mutase , chemistry , mass spectrometry , enzyme , ammonium bicarbonate , biochemistry , phosphorylation , electrospray , hydrolysis , chromatography , phosphoglycerate kinase , glycolysis , organic chemistry , raw material
Electrospray mass spectrometry has been used to study the formation and hydrolysis of the phosphorylated forms of two phosphoglycerate mutases. The half‐life of the enzyme from Saccharomyces cerevisiae was 35 min at 20°C in 10 mM ammonium bicarbonate, pH 8.0. Addition of 1 mM 2‐phosphoglycollate reduced this value by at least 100‐fold. The phosphorylated form of the enzyme from Schizosaccharomyces pombe was much less stable with a half‐life of less than 1 min. The results are discussed in terms of the kinetic properties of the enzymes. Mass spectrometry would appear to be a powerful method to study the formation and breakdown of phosphorylated proteins, processes which are of widespread significance in regulatory mechanisms.