Effects on Ca(2+)-activated tension due to a synthetic NH2-terminal actin peptide in single skeletal muscle fibers.
Insight into the mechanism of force development in striated muscle will be provided by elucidating the specific regions of the actin molecule that interact with myosin and regulatory subunits of the thin filament during Ca(2+)-activated contraction. There is growing evidence that the acidic NH2-terminal domain of actin 1) may represent an important binding site for myosin and 2) may interact with the inhibitory region of troponin I. The purpose of this study was to determine the effects of a synthetic peptide corresponding to a specific sequence of the NH2-terminal domain of skeletal muscle actin on Ca(2+)-activated tension in chemically skinned single psoas skeletal muscle fibers. This study focused on the highly conserved Lys18-Arg28 amino acid sequence of actin, a region of native actin that is believed to interact with troponin I and myosin. The effects of synthetic actin peptide Lys18-Arg28 on tension development varied, depending on 1) the concentration of Ca2+ in the activating solutions and 2) the peptide concentration. At submaximal concentrations of Ca2+, isometric tension was reversibly potentiated in the presence of 100-500 microM synthetic actin peptide Lys18-Arg28. Importantly, scrambling the sequence of Lys18-Arg28 fully abolished the increase in Ca2+ sensitivity, providing evidence that the observed effects were specific to the sequence of peptide Lys18-Arg28. In contrast, maximum Ca(2+)-activated tension was inhibited by millimolar concentrations of Lys18-Arg28 and the scrambled peptide, indicating that this effect was nonspecific. The effect of peptide Lys18-Arg28 to increase the Ca2+ sensitivity of tension is not known but may be due to an effect of the actin peptide to alter thin filament activation, a possibility consistent with proposed interactions between this domain of actin and the inhibitory region of troponin I.
D000199:Actins / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000502:physiology*; D066298:In Vitro Techniques; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects*; D018485:Muscle Fibers, Skeletal / Q000187:drug effects*; D018482:Muscle, Skeletal / Q000187:drug effects*; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000494:pharmacology*; D011817:Rabbits
D000199:Actins; D010446:Peptide Fragments; D002118:Calcium