Epidermal growth factor receptor tyrosine kinase phosphorylation of glucose-6-phosphate dehydrogenase in vitro.
The specific tyrosine phosphorylation of glucose-6-phosphate dehydrogenase (G6PDH) by the epidermal growth factor (EGF) receptor in vitro is demonstrated. The Km values of the substrate G6PDH and of ATP for the receptor tyrosine kinase were ca. 1 and 10 microM, respectively. The rate of phosphorylation was EGF dependent, with a four-fold increase in Vmax in the presence of EGF. The phosphorylation was stimulated maximally by 0.2 microM or greater EGF, with an ED50 of ca. 20 nM which is consistent with the affinity of the solubilized receptor for EGF. Using conditions of 5 microM G6PDH, 100 microM ATP, 5 mM Mg2+, and 1 mM Mn2+, up to 0.3 mol phosphate was incorporated into 1 mol of the 55-kDa subunit of Baker's yeast G6PDH. Tryptic peptide mapping revealed several unique phosphopeptides for both Baker's yeast and bovine adrenal G6PDH. The patterns of phosphopeptides for a given enzyme were identical for basal and EGF-stimulated phosphorylation.
D000818:Animals; D002417:Cattle; D003454:Cryptococcus / Q000201:enzymology; D066246:ErbB Receptors / Q000378:metabolism*; D001411:Geobacillus stearothermophilus / Q000201:enzymology; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D007934:Leuconostoc / Q000201:enzymology; D008274:Magnesium / Q000378:metabolism; D008345:Manganese / Q000378:metabolism; D015688:Oncogene Protein pp60(v-src); D010449:Peptide Mapping; D010750:Phosphoproteins / Q000032:analysis; D010751:Phosphopyruvate Hydratase / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011817:Rabbits; D012191:Retroviridae Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology
D010750:Phosphoproteins; D012191:Retroviridae Proteins; D008345:Manganese; D005954:Glucosephosphate Dehydrogenase; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src); D010751:Phosphopyruvate Hydratase; D008274:Magnesium