Open AccessSaccharomyces cerevisiae pyruvate kinase Pyk1 is PKA phosphorylation substrate in vitro
Author(s) -
Cytryńska Małgorzata,
Frajnt Magdalena,
Jakubowicz Teresa
Publication year - 2001
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2001.tb10845.x
Subject(s) - protein kinase a , saccharomyces cerevisiae , biochemistry , protein subunit , phosphorylation , yeast , in vitro , chemistry , pyruvate kinase , enzyme , biology , glycolysis , gene
Fractionation of Saccharomyces cerevisiae postribosomal extract on DEAE‐cellulose revealed two fractions of cAMP‐dependent protein kinase (PKA‐1 and PKA‐2). The presence of PKA in both fractions was confirmed by immunoblotting with anti‐Bcy1 antibodies. Yeast pyruvate kinase Pyk1 identified by amino acid microsequencing analysis and immunoblotting with anti‐Pyk1 antibodies copurified with the PKA‐1 but not the ‐2 fraction. Pyk1 can be phosphorylated by yeast PKA in vitro in the presence of cAMP and cGMP. Two‐dimensional gel electrophoretic analysis revealed four phosphorylated forms of Pyk1 modified by PKA. In phosphorylation of Pyk1 mainly the Tpk2 catalytic subunit of yeast PKA was involved.