Effect of negative mechanical stress on the orientation of myosin cross-bridges in muscle fibers.
The effect of positive and negative stress on myosin cross-bridge orientation in glycerinated muscle fibers was investigated by using fluorescence polarization spectroscopy of the emission from the covalent label tetramethyl-rhodamine-5-(and -6)-iodoacetamide (IATR) specifically modifying sulfhydryl one (SH1) on the myosin heavy chain. Positive tension was applied by stretching the fiber in rigor. Negative tension was applied in two steps by using a protocol introduced by Goldman et al. [Goldman, Y. E., McCray, J. A. & Vallette, D. P. (1988) J. Physiol. (London) 398, 75P]: relaxing a fiber at resting length and stretching it until the relaxed tension is appreciable and then placing the fiber in rigor and releasing the tension onto the rigor cross-bridges. We found, as have others, that positive tension has no effect on the fluorescence polarization spectrum from the SH1-bound probe, indicating that the cross-bridge does not rotate under these conditions. Negative tension, however, causes a change in the fluorescence polarization spectrum that indicates a probe rotation. The changes in the polarization spectrum from negative stress are partially reversed by the subsequent application of positive stress. It appears that negative tension strains the cross-bridge, or the cross-bridge domain containing SH1, and causes it to rotate.
Burghardt
T P
TP
Department of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, MN 55905.
Ajtai
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D000199:Actins / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D046911:Macromolecular Substances; D008433:Mathematics; D009132:Muscles / Q000502:physiology*; D009218:Myosins / Q000378:metabolism*; D010446:Peptide Fragments / Q000032:analysis; D011817:Rabbits; D013050:Spectrometry, Fluorescence; D013314:Stress, Mechanical
D000199:Actins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D000251:Adenosine Triphosphatases; D009218:Myosins