Manganese modulates protein phosphorylation in the rat pancreas: In vitro evidence for cation selective regulation

J. Scott Brockenbrough, Murray Korc

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

The effects of manganese (Mn2+) on phosphorylation activity in the rat pancreas were examined in an in vitro phosphorylation assay and by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of endogenous proteins. Several histones were phosphorylated in the presence of pancreatic supernatant obtained following a 10-min centrifugation at 15,000 g. The histone preference for this reaction was VII-S > V-S or II-S > III-S > other histones. The Mn2+(10 mM) enhanced the phosphorylation of some histones (II-A, III-S, VI-S, and VIII-S) but inhibited the phosphorylation of other histones (II-S, V-S, and VII-S). The same concentration of Mn2+also enhanced the incorporation of32P into cytosolic proteins in the absence of exogenous histones. This effect was not mimicked by Ca2+, Mg2+, Ba2+, or Zn2+. Analysis of endogenous proteins by SDS-PAGE revealed Mn2+-dependent and time-dependent phosphorylation of high (98-200-kDa), intermediate (59-,52-,35-, and 30-kDa), and low-molecular weight proteins. The Mn2+exerted similar effects in the presence of pancreatic cytosol obtained following a 60-min centrifugation at 100,000 g. However, the 35-and 30-kDa phosphoproteins and the low-molecular weight proteins were not readily visible. In both the 15,000-and the 100,000-g preparations, there was a dose-dependent increase in the phosphorylation of the Mr 98-kDa protein (p98) at concentrations ranging from 0.03 to 1.0 mM Mn2+but a lesser stimulatory effect at 10.0 mM Mn2+. The effects of Mn2+on the phosphorylation of specific pancreatic proteins were not mimicked by other divalent cations. These findings suggest that the rat exocrine pancreas may possess a specific Mn2+-dependent kinase.

Original languageEnglish (US)
Pages (from-to)589-597
Number of pages9
JournalPancreas
Volume5
Issue number5
DOIs
StatePublished - Sep 1990

Keywords

  • Kinase activity
  • Manganese
  • Pancreas
  • Phosphorylation

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology

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