Differential autophosphorylation of CaM kinase II from phasic and tonic smooth muscle tissues.
Ca+/calmodulin-dependent protein kinase II (CaM kinase II) is regulated by calcium oscillations, autophosphorylation, and its subunit composition. All four subunit isoforms were detected in gastric fundus and proximal colon smooth muscles by RT-PCR, but only the gamma and delta isoforms are expressed in myocytes. Relative gamma and delta message levels were quantitated by real-time PCR. CaM kinase II protein and Ca2+/calmodulin-stimulated (total) activity levels are higher in proximal colon smooth muscle lysates than in fundus lysates, but Ca2+/calmodulin-independent (autonomous) activity is higher in fundus lysates. CaM kinase II in fundus lysates is relatively unresponsive to Ca2+/calmodulin. Alkaline phosphatase decreased CaM kinase II autonomous activity in fundus lysates and restored its responsiveness to Ca2+/calmodulin. Acetylcholine (ACh) increased autonomous CaM kinase II activity in fundus and proximal colon smooth muscles in a time- and dose-dependent manner. KN-93 enhanced ACh-induced fundus contractions but inhibited proximal colon contractions. The different properties of CaM kinase II from fundus and proximal colon smooth muscles suggest differential regulation of its autophosphorylation and activity in tonic and phasic gastrointestinal smooth muscles.
Lorenz
Jillinda M
JM
Department of Physiology and Cell Biology, University of Nevada School of Medicine, Reno, Nevada 89557, USA.
Riddervold
Marilyn H
MH
Beckett
Elizabeth A H
EA
Baker
Salah A
SA
Perrino
Brian A
BA
D000109:Acetylcholine / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D017398:Alternative Splicing / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D001596:Benzylamines / Q000494:pharmacology; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000235:genetics* / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism; D003106:Colon / Q000201:enzymology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005748:Gastric Fundus / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009130:Muscle, Smooth / Q000201:enzymology*; D010766:Phosphorylation; D013449:Sulfonamides / Q000494:pharmacology; D000096926:Benzenesulfonamides
D000109:Acetylcholine; D000469:Alkaline Phosphatase; D001596:Benzylamines; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002147:Calmodulin; D004791:Enzyme Inhibitors; D013449:Sulfonamides; C072105:KN 93; D000096926:Benzenesulfonamides