Myofibrillar calcium sensitivity modulation: influence of light chain phosphorylation and positive inotropic drugs on skinned frog skeletal muscle.
The myofibrillar calcium sensitivity of mammalian skeletal muscle and cardiac muscle may be increased by myosin light chain kinase (MLCK)-induced myosin phosphorylation5) 13). Here we report increasing calcium responsiveness of frog skeletal muscle fibres (Tibialis anterior, skinned by freeze drying) by MLCK-induced myosin P-light chain phosphorylation and by the non-glycoside, non-sympathomimetic positive inotropic drug pimobendan. Investigation of myosin light chains by two dimensional gel electrophoresis revealed two phosphorylatable P-light chains (LC-2) having the same isoelectric point (5.3 for the unphosphorylated, 5.1 for the phosphorylated form) but different molecular weights (19 and 18 kD, respectively). This pattern of LC-2 is distinct from mammalian skeletal and cardiac muscle (only one phosphorylatable P-light chain in skeletal, two phosphorylatable P-light chains in cardiac muscle with different isoelectric points, but identical molecular weight). The phosphorylation level was about 0.45 mole phosphate/mole P-light chains and could be increased by 16 +/- 3% by the addition of myosin light chain kinase. This procedure increased the isometric tension at pCa 5.5 by 21 +/- 5% while maximum tension (at pCa 4.3) was not affected by MLCK. The new inotropic drug pimobendan (10(-4) M) increased isometric tension at pCa 6 by 48 +/- 4.5%, but maximal tension was not affected. Another positive inotropic drug, sulmazole, has been shown to potentiate the twitch of intact frog Tibialis anterior muscle23) and to increase force of skinned fibres by 21.7 +/- 3.3% at submaximal activation (pCa 6).
Morano
I
I
II. Physiologisches Institut, Universität Heidelberg, Federal Republic of Germany.
Piazzesi
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G
Rüegg
J C
JC
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002316:Cardiotonic Agents / Q000494:pharmacology*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D009210:Myofibrils / Q000187:drug effects / Q000502:physiology*; D015879:Myosin Subfragments; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D010766:Phosphorylation; D011724:Pyridazines / Q000494:pharmacology*; D011893:Rana esculenta
D002316:Cardiotonic Agents; D007093:Imidazoles; D015879:Myosin Subfragments; D010446:Peptide Fragments; D011724:Pyridazines; C041648:pimobendan; D009219:Myosin-Light-Chain Kinase; D009218:Myosins; C029758:sulmazole; D002118:Calcium