Characterization of gamma- and delta-subunits of Ca2+/calmodulin-dependent protein kinase II in rat gastric mucosal cell populations.
We searched for the occurrence of a Ca2+/calmodulin-dependent protein kinase in rat gastric cell types as a likely member in the chain of gastrin- and muscarinic-receptor-mediated signal transmission. A Ca(2+)- and calmodulin-dependent phosphorylation of major 50, 60 and 100 kDa substrates was observed in parietal cell cytosol and a major 60 and 61 kDa protein doublet was found to bind 125I-calmodulin in 125I-calmodulin-gel overlays. A specific substrate of the multifunctional Ca2+/calmodulin-dependent protein kinase II, autocamtide II, was phosphorylated in a calmodulin-dependent manner. The specific inhibitor of this enzyme, KN-62, antagonized protein kinase activity. RNA extracted from gastric mucosal cells was shown to contain sequences of the gamma- and delta- but not alpha- and beta-subunits of the calmodulin-dependent kinase II, and mRNA of both subtypes was demonstrated in highly purified parietal, chief and mucous cells. A calmodulin-dependent kinase II composed of gamma- and delta-subunits is a likely mediator of Ca(2+)-dependent signal transmission in these populations of gastric cells.
Mayer
P
P
Medizinische Klinik und Poliklinik, Berufsgenossenschaftliche Krankenanstalten Bergmannsheil, Rurh-Universität, Bochum, Germany.
Möhlig
M
M
Seidler
U
U
Rochlitz
H
H
Fährmann
M
M
Schatz
H
H
Hidaka
H
H
Pfeiffer
A
A
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives*; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002118:Calcium / Q000494:pharmacology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D002147:Calmodulin / Q000494:pharmacology; D005753:Gastric Mucosa / Q000166:cytology / Q000201:enzymology*; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity
D000911:Antibodies, Monoclonal; D002147:Calmodulin; D007546:Isoquinolines; D010879:Piperazines; D012333:RNA, Messenger; C063302:KN 62; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002118:Calcium