Creatine kinase activity associated with the contractile proteins of the guinea-pig carotid artery.
Activity and role of creatine kinase associated with contractile proteins of vascular smooth muscle have been investigated using skinned guinea-pig carotid artery rings. Membrane solubilization was performed with the detergent Triton X-100. Creatine kinase activity, isoenzyme profile as well as mechanics were performed on the Triton skinned carotid artery rings. Total creatine kinase activity was 47.3 +/- 9.3 IU g-1 ww and electrophoresis showed BB, MB, and MM isoforms (BB-CK being the predominant isoenzyme). One hour incubation with Triton X-100, produced predominantly BB-CK remaining with the myofibrils with some MB, representing 23% of the preskinned creatine kinase activity. When relaxed carotid artery rings were exposed to pCa 9 in the presence of 250 microM ADP, 0 ATP, and 12 mM phosphocreatine, tension was not significantly different from resting tension, but changing to pCa 4.5 caused the carotid artery rings to generate 49.5 +/- 4.5% of maximal tension. When a high-tension rigor state was achieved (250 microM ADP, 0 ATP, 0 phosphocreatine, and pCa 9), the addition of 12 mM phosphocreatine effected significant relaxation. These observations implicate an endogenous form of creatine kinase, associated with the myofilaments, which is capable of producing enough ATP for submaximal tension generation and significant relaxation from rigor conditions. These results suggest co-localization of ATPase, MLCK, and creatine kinase on the contractile proteins of the carotid artery. Such an enzymic association may play a role in the energetic supply to the contractile apparatus of vascular smooth muscle.
Clark
J F
JF
Department of Biochemistry, University of Oxford, UK.
Khuchua
Z
Z
Boehm
E
E
Ventura-Clapier
R
R
D000244:Adenosine Diphosphate / Q000096:biosynthesis; D000818:Animals; D002339:Carotid Arteries / Q000378:metabolism*; D003402:Creatine Kinase / Q000096:biosynthesis*; D006168:Guinea Pigs; D009119:Muscle Contraction; D009131:Muscle, Smooth, Vascular / Q000201:enzymology* / Q000502:physiology; D009218:Myosins / Q000096:biosynthesis
D000244:Adenosine Diphosphate; D003402:Creatine Kinase; D009218:Myosins