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Małgorzata Cytryńska, Magdalena Frajnt, Teresa Jakubowicz, Saccharomyces cerevisiae pyruvate kinase Pyk1 is PKA phosphorylation substrate in vitro, FEMS Microbiology Letters, Volume 203, Issue 2, September 2001, Pages 223–227, https://doi.org/10.1111/j.1574-6968.2001.tb10845.x
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Abstract
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.