title: Purification and characterization of calcium-calmodulin kinase II from human parathyroid glands.
abstract: Calmodulin has been identified in parathyroid cells and is thought to play an important role in the production or secretion of parathyroid hormone. However, a detailed investigation of calmodulin-binding proteins in parathyroid glands has not been conducted. In this study, we attempted to determine the presence of calmodulin-binding protein in human parathyroid adenoma by affinity chromatography. The eluted protein from a calmodulin-coupled Sepharose 4B column with EGTA was analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis which revealed a major protein band of Mr 50,000. A Ca2+/calmodulin-dependent protein kinase activity was detected at the protein peak using dephosphorylated casein as a substrate. The 50 kDa band was identified as calcium/calmodulin-dependent protein kinase II (CaM-kinase II) by immunoblotting. The substrate specificity, pH dependency and affinity for calmodulin of this enzyme were identical to those of CaM-kinase II from rat brain. Also, the kinase activity was sensitive to KN-62, a specific inhibitor of CaM-kinase II. In total, 0.48 mg of this kinase was purified from 3 g human parathyroid adenoma.
authors.affiliation: Department of Pharmacology, Nagoya University School of Medicine, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kato
authors.forename: M
authors.initials: M
authors.lastname: Hagiwara
authors.forename: Y
authors.initials: Y
authors.lastname: Nimura
authors.forename: S
authors.initials: S
authors.lastname: Shionoya
authors.forename: H
authors.initials: H
authors.lastname: Hidaka
mesh_terms: D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D015151:Immunoblotting; D010280:Parathyroid Glands / Q000737:chemistry*; D010766:Phosphorylation; D011494:Protein Kinases / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases