Cloning and quantitative determination of the human Ca2+/calmodulin-dependent protein kinase II (CaMK II) isoforms in human beta cells

被引:22
作者
Rochlitz, H
Voigt, A
Lankat-Buttgereit, B
Göke, B
Heimberg, H
Nauck, MA
Schiemann, U
Schatz, H
Pfeiffer, AFH
机构
[1] Ruhr Univ Bochum, Hosp Bergmannsheil, Dept Internal Med, Bochum, Germany
[2] Univ Marburg, Dept Internal Med, Clin Res Unit Gastrointestinal Endocrinol, D-3550 Marburg, Germany
[3] Free Univ Brussels, Diabet Res Ctr, Brussels, Belgium
[4] Ruhr Univ Bochum, Knappsch Hosp, Dept Internal Med, Bochum, Germany
[5] Univ Munster Hosp, Dept Internal Med B, Munster, Germany
关键词
insulin secretion; protein kinase; insulin secreting cells; human CaMK II; cloning of new subtypes;
D O I
10.1007/s001250051330
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Aims/hypothesis. The Ca2+/calmodulin-dependent protein kinase II (CaMK II) is highly expressed in pancreatic islets and associated with insulin secretion vesicles. The suppression of CaMK II disturbs insulin secretion and insulin gene expression. There are four isoforms of CaMK II, alpha to delta, that are expressed from different genes in mammals. Our aim was to identify the isoforms of CaMK II expressed in human beta cells by molecular cloning from a human insulinoma cDNA library and to assess its distribution in humans. Methods. The previously unknown complete coding sequences of human CaMK II beta and the kinase domain of CaMK II delta were cloned from a human insulinoma cDNA library. Quantitative determination of CaMK II isoform mRNA was carried out in several tissues and beta cells purified by fluorescence activated cell sorting and compared to the housekeeping enzyme pyruvate dehydrogenase. Results. We found CaMK II beta occurred in three spice variants and was highly expressed in endocrine tissues such as adrenals, pituitary and beta cells. Liver showed moderate expression but adipose tissue or lymphocytes had very low levels of CaMK II beta-mRNA. In human beta cells CaMK II beta and delta were expressed equally with pyruvate dehydrogenase whereas tenfold lower expression of CaMK II gamma and no expression of CaMK II alpha were found. Conclusion/interpretation. Although CaMK II delta is ubiquitously expressed, CaMK II beta shows preferential expression in neuroendocrine tissues. In comparison with the expression of a key regulatory enzyme in glucose oxidation, pyruvate dehydrogenase, two of the four CaM kinases investigated are expressed at equally high levels, which supports an important role in beta-cell physiology. These results provide the basis for exploring the pathophysiological relevance of CaMK II beta in human diabetes.
引用
收藏
页码:465 / 473
页数:9
相关论文
共 51 条
[31]   INTERACTION OF AUTOPHOSPHORYLATED CA2+ CALMODULIN-DEPENDENT PROTEIN-KINASE-II WITH NEURONAL CYTOSKELETAL PROTEINS - CHARACTERIZATION OF BINDING TO A 190-KDA POSTSYNAPTIC DENSITY PROTEIN [J].
MCNEILL, RB ;
COLBRAN, RJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (17) :10043-10049
[32]   Insulinoma cells contain an isoform of Ca2+/calmodulin-dependent protein kinase II delta associated with insulin secretion vesicles [J].
Mohlig, M ;
Wolter, S ;
Mayer, P ;
Lang, J ;
Osterhoff, M ;
Horn, PA ;
Schatz, H ;
Pfeiffer, A .
ENDOCRINOLOGY, 1997, 138 (06) :2577-2584
[33]  
NGHIEM P, 1993, J BIOL CHEM, V268, P5471
[34]   PRESENCE AND POSSIBLE INVOLVEMENT OF CA/CALMODULIN-DEPENDENT PROTEIN-KINASES IN INSULIN RELEASE FROM THE RAT PANCREATIC BETA-CELL [J].
NIKI, I ;
OKAZAKI, K ;
SAITOH, M ;
NIKI, A ;
NIKI, H ;
TAMAGAWA, T ;
IGUCHI, A ;
HIDAKA, H .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1993, 191 (01) :255-261
[35]  
Osterhoff M, 1999, DIABETOLOGIA, V42, pA25
[36]  
Pfeiffer D, 1997, INT J CANCER, V72, P581, DOI 10.1002/(SICI)1097-0215(19970807)72:4<581::AID-IJC5>3.0.CO
[37]  
2-M
[38]   PHYSIOLOGY AND PATHOPHYSIOLOGY OF INSULIN-SECRETION [J].
RASMUSSEN, H ;
ZAWALICH, KC ;
GANESAN, S ;
CALLE, R ;
ZAWALICH, WS .
DIABETES CARE, 1990, 13 (06) :655-666
[39]   MULTIFUNCTIONAL CA-2+/CALMODULIN-DEPENDENT PROTEIN-KINASE - DOMAIN-STRUCTURE AND REGULATION [J].
SCHULMAN, H ;
LOU, LL .
TRENDS IN BIOCHEMICAL SCIENCES, 1989, 14 (02) :62-66
[40]   MULTIFUNCTIONAL CA2+/CALMODULIN-DEPENDENT PROTEIN-KINASE [J].
SCHULMAN, H ;
HANSON, PI .
NEUROCHEMICAL RESEARCH, 1993, 18 (01) :65-77