Identification of Raf‐1 Ser<sup>621</sup> kinase activity from NIH 3T3 cells as AMP‐activated protein kinase
FEBS LettersPublished 24 February 1997Open access
Amy B. Sprenkle, Stephen Davies, David Carling, D. Grahame Hardie, Thomas W. Sturgill
Citations62
SJR quartileQ1
SJR score1.22
SNIP0.77
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This poster presents a probabilistic procedure for determining the phytochemical properties of polymethine and its role in the response to infectious disease and its applications in medicine.
Abstract
Raf-1 is extensively phosphorylated on Ser621 in both quiescent and mitogen-stimulated cells. To identify the responsible kinase(s), cytosolic fractions of NIH 3T3 cells were analyzed for Ser621 peptide kinase activity. One major peak of activity was detected and identified as AMP-activated protein kinase (AMPK) by immunodepletion experiments. AMPK phosphorylated the catalytic domain of Raf-1, expressed in Escherichia coli as a soluble GST fusion protein, to generate a single tryptic [32P]phosphopeptide containing exclusively phospho-Ser621. AMPK also phosphorylated full-length, kinase-defective Raf-1 (K375M) to generate two [32P]phosphopeptides, one co-migrating with synthetic tryptic peptide containing phospho-Ser621 and the other with phospho-Ser259.
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Biochemistry, Genetics and Molecular Biology
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