title: Polyamines regulate serine/threonine protein phosphatases in insulin-secreting cells.
abstract: Reversible protein phosphorylation is an important mechanism by which cells transduce external signals into biologic responses. Levels of protein phosphorylation are determined by the balanced actions of both protein kinases and protein phosphatases (PPases). However, compared with protein kinases, regulation of PPases has been relatively neglected. The insulin secretagogue L-arginine, an immediate metabolic precursor to polyamines, causes a rapid and transient decrease in PPase-1 activity in insulin-secreting RINm5F cells. We here show that polyamines dose-dependently suppress PPase-1-like activity when added to RINm5F cell homogenates at physiologic concentrations (spermine > spermidine > putrescine), while having minor and inconsistent effects on PPase-2A-like activity. The IC50 value for spermine on PPase-1-like activity was approximately 4 mM. The inhibitory effect was reproduced and of comparable magnitude on purified PPases types 1 and 2A. On the other hand, when endogenous polyamine pools were exhausted by 4 days of exposure to the specific L-ornithine decarboxylase inhibitor DL-alpha-difluoromethylornithine, there was an increase in PPase-2A-like activity. Quantitative Western analysis revealed that the amount of PPase-2A protein did not change after this treatment. It is concluded that polyamines cause time-and concentration-dependent inhibitory effects on RINm5F cell PPase activities, which may contribute to the increase in phosphorylation state that occurs after secretory stimulation.
authors.affiliation: Department of Molecular Medicine, The Rolf Luft Center for Diabetes Research, Karolinska Institute, Karolinska Hospital, Stockholm, Sweden. ake@enk.ks.se
authors.forename: A
authors.initials: A
authors.lastname: Sjöholm
authors.forename: R E
authors.initials: RE
authors.lastname: Honkanen
mesh_terms: D001120:Arginine / Q000378:metabolism; D000518:Eflornithine / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007340:Insulinoma / Q000378:metabolism / Q000473:pathology; D007515:Islets of Langerhans / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D019319:Okadaic Acid / Q000494:pharmacology; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D011073:Polyamines / Q000494:pharmacology*; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011700:Putrescine / Q000494:pharmacology; D013095:Spermidine / Q000494:pharmacology; D013096:Spermine / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D004791:Enzyme Inhibitors; D006657:Histones; D007328:Insulin; D011073:Polyamines; D019319:Okadaic Acid; D013096:Spermine; D001120:Arginine; D017346:Protein Serine-Threonine Kinases; C067024:okadaic acid-sensitive phosphatase; D010749:Phosphoprotein Phosphatases; D013095:Spermidine; D011700:Putrescine; D000518:Eflornithine