Polyamine-sensitive protein kinase from chick intestinal mucosa.
A cyclic nucleotide-independent protein kinase which phosphorylates preferentially acidic proteins such as casein or phosvitin was isolated from cytosol of chick duodenal mucosa. The enzyme was purified more than 633 fold to apparent homogeneity by ammonium sulfate fractionation, column chromatography on DEAE-cellulose, phosphocellulose, hydroxylapatite and by sucrose density gradient centrifugation. The native enzyme has a molecular weight of 131 000 as measured by gel filtration. The enzyme is a complex protein containing three polypeptides of molecular weight of 39 000, 36 000 and 27 000. It behaves as a complex throughout its purification and gel filtration but its components are readily separated by electrophoresis in denaturing buffer. The 27 000 molecular weight band was selectively autophosphorylated when the enzyme was incubated in the presence of [gamma-32P]ATP. When casein was used as substrate, physiological concentrations of naturally occurring polyamines such as spermine and spermidine markedly stimulated enzyme activity. However with phosvitin as substrate polyamines were strong inhibitors of the enzyme activity. This contrasting effect on intestinal kinase activity was also apparent using cytoplasmic proteins as endogenous phosphate acceptors. A characterization of this differential effect is presented and some possible physiological implications are discussed.
D000818:Animals; D001345:Autoradiography; D002364:Caseins / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D002645:Chickens; D002845:Chromatography / Q000379:methods; D003593:Cytoplasm / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation / Q000187:drug effects; D007413:Intestinal Mucosa / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D010774:Phosvitin / Q000378:metabolism; D011073:Polyamines / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism
D002364:Caseins; D011073:Polyamines; D047428:Protein Kinase Inhibitors; D010774:Phosvitin; D011494:Protein Kinases