Phosphorylation of gastrin-related peptides: physiological casein kinase like enzyme in Golgi membranes from bovine adrenal chromaffin cells and GH3 cells.
Prohormones such as the gastrin precursor can be phosphorylated at Ser residues, on passage along the secretory pathway. The phosphorylation site occurs in a sequence (-Ser-Ala-Glu-) that suggests these peptides are substrates for physiological casein kinase, but the presence of this enzyme in endocrine cells is unknown. We have examined the specificity of Golgi membrane kinases from lactating rat mammary gland, bovine adrenal medulla and the GH3 cell line, for phosphorylation of progastrin fragments and analogues. The kinetics of phosphorylation of peptides with the native sequence, -Arg-Arg-Ser-Ala-Glu- were similar to those of tryptic cleavage fragments (Ser-Ala-Glu-) in both mammary and endocrine cell preparations. The product of in vitro phosphorylation was chromatographically indistinguishable from native peptide. Peptides with the sequence Ser-Ala-Ala (i.e., substitution of Glu to Ala) were not phosphorylated. We conclude that a physiological casein kinase like enzyme can act on both the gastrin precursor and its COOH-terminal cleavage product, and occurs in the Golgi complex of both mammary gland and peptide-producing endocrine cells.
Vegh
M
M
Physiological Laboratory, University of Liverpool, UK.
Varro
A
A
D000255:Adenosine Triphosphate; D000313:Adrenal Medulla / Q000201:enzymology; D000409:Alanine; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium; D047388:Casein Kinases; D002413:Cations, Divalent; D002417:Cattle; D002462:Cell Membrane; D019439:Chromaffin Cells / Q000166:cytology / Q000201:enzymology; D005260:Female; D005755:Gastrins / Q000378:metabolism*; D018698:Glutamic Acid; D006056:Golgi Apparatus / Q000201:enzymology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008274:Magnesium; D008321:Mammary Glands, Animal; D008345:Manganese; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured
D002413:Cations, Divalent; D005755:Gastrins; D010455:Peptides; D018698:Glutamic Acid; D008345:Manganese; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D047388:Casein Kinases; D008274:Magnesium; D000409:Alanine; D002118:Calcium