title: Human islets of Langerhans express multiple isoforms of calcium/calmodulin-dependent protein kinase II.
abstract: Previous studies have provided evidence for the presence of calcium/calmodulin-dependent protein kinase II (CaM kinase II) in rodent islets of Langerhans, and beta-cell CaM kinase II activity has been correlated with insulin secretion. In this study we provide the first conclusive evidence for the expression of CaM kinase II in human islets of Langerhans and show that multiple isoforms are expressed. Screening of a human islet cDNA library resulted in the isolation of a 999bp partial cDNA clone encoding CaM kinase II. The nucleotide sequence of the islet clone showed a high degree of homology (94.8%) to the two gamma isoforms of CaM kinase II previously isolated from human T lymphocytes (gammaB and gammaC). In order to obtain full length sequence for the islet clone, rapid amplification of cDNA ends (RACE) was used to amplify the 3' end of the islet clone from human islet poly A+ RNA. Two distinct gamma isoforms of CaM kinase II were amplified from the islet RNA. They were identified as gammaB and gammaE; the latter is distinguished from gammaB by a 114bp insertion within the association domain of the cDNA. Using reverse transcriptase polymerase chain reaction (RT-PCR) we also detected in human islets of Langerhans the novel beta3 isoform of CaM kinase II previously reported to be expressed in neonatal rat islets.
authors.affiliation: Nuffield Department of Clinical Biochemistry, John Radcliffe Hospital, Headington, Oxford, United Kingdom.
authors.forename: M A
authors.initials: MA
authors.lastname: Breen
authors.forename: S J
authors.initials: SJ
authors.lastname: Ashcroft
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D006801:Humans; D007515:Islets of Langerhans / Q000201:enzymology*; D007527:Isoenzymes / Q000378:metabolism*; D008969:Molecular Sequence Data
chemical_list: D007527:Isoenzymes; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases