Identification of the calmodulin-regulated protein phosphatase, calcineurin, in rat pancreatic islets.
The aim of this work was the identification of the calmodulin-stimulated protein phosphatase, calcineurin, in rat pancreatic islets. For this purpose, a high-affinity calcineurin antibody and the Western blotting technique were used to detect the presence of calcineurin in freshly collagenase-isolated islets. The calcineurin content detected by this method was about 0.30 ng islet (approx. 0.07% of the total islet protein). The subunit composition and Mr of islet calcineurin were similar to those of bovine brain calcineurin. Incubation of nitrocellulose membranes of the Western blotting, containing the islet protein fractions, with 125I-labeled calmodulin and 45Ca2+ demonstrated that the A subunit bound calmodulin, while the B subunit bound Ca2+. The presence of calcineurin in the islets of Langerhans would suggest its possible participation, as a counterpart of the kinases effect, in the regulatory mechanism of insulin secretion.
Gagliardino
J J
JJ
CENEXA-Centro de Endocrinología Experimental y Aplicada (UNLP-CONICET), Facultad de Ciencas Médicas, La Plata, Argentina.
Krinks
M H
MH
Gagliardino
E E
EE
D000818:Animals; D015153:Blotting, Western; D019703:Calcineurin; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D007515:Islets of Langerhans / Q000201:enzymology*; D046911:Macromolecular Substances; D008297:Male; D008970:Molecular Weight; D010749:Phosphoprotein Phosphatases / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D046911:Macromolecular Substances; D019703:Calcineurin; D010749:Phosphoprotein Phosphatases; D002118:Calcium