title: Isolation, purification and cell-free synthesis of calmodulin from the pig anterior pituitary gland.
abstract: Calmodulin was extracted and purified from pig anterior pituitary gland. The protein was characterized by its migration on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis in the presence of Ca2+ or EGTA, its U.V. spectrum between 240 and 290 nm and the activation of calmodulin-deficient cyclic AMP phosphodiesterase. The yield was 370 mg/kg wet wt. mRNA was also extracted from the same tissue and translated in a wheat-germ cell-free translation system. Translated calmodulin was identified by its heat-stability, its co-migration with authentic anterior-pituitary calmodulin on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, its acidic isoelectric point (4.15) on flat-bed isoelectric focusing, its Ca2+-dependent binding to fluphenazine-Sepharose 6B, and its co-elution from this gel with authentic unlabelled calmodulin with EGTA buffer. Calmodulin was not translated as a precursor form. In this tissue it was calculated that calmodulin accounted for 0.5-1% of the total translated protein.
mesh_terms: D000818:Animals; D002147:Calmodulin / Q000096:biosynthesis / Q000302:isolation & purification*; D002474:Cell-Free System; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D010903:Pituitary Gland, Anterior / Q000032:analysis*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine
chemical_list: D002147:Calmodulin; D012333:RNA, Messenger