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Biophys J, August 1998, p. 948-956, Vol. 75, No. 2
*Department of Physiology and Neurobiology, University of Connecticut, Storrs, Connecticut 06269 and #Sloan-Kettering Research Center, New York, New York 10024 USA
Measurements were made of the distributions of total
calcium along the length of A and I bands in skinned frog
semitendinosus muscles using electron probe x-ray microanalysis. Since
calcium in the water space was kept below the detection limit of the
technique, the signal was assumed to reflect the distribution of
calcium bound to myofilament proteins. Data from sarcomeres with
overlap between thick and thin filaments showed enhancement of calcium in this region, as previously demonstrated in rabbit psoas muscle fibers in rigor (Cantino, M. E., T. S. Allen, and A. M. Gordon. 1993. Subsarcomeric distribution of calcium in demembranated
fibers of rabbit psoas muscle. Biophys. J. 64:211-222).
Such enhancement could arise from intrinsic non-uniformities in calcium
binding to either thick or thin filaments or from enhancement of
calcium binding to either filament by rigor cross-bridge attachment. To test for intrinsic variations in calcium binding, calcium distributions were determined in fibers stretched to beyond filament overlap. Calcium
binding was found to be relatively uniform along both thick and thin
filaments, and therefore cannot account for the increased calcium
observed in the overlap region. From these results it can be concluded
that the observed enhancement of calcium is due to an increase in
calcium binding to myofilaments as a result of rigor attachment of
cross-bridges to actin. The source of the enhancement is most likely an
increase in calcium binding to troponin, although enhancement of
calcium binding to myosin light chains cannot be ruled out.
Biophys J, August 1998, p. 948-956, Vol. 75, No. 2
© 1998 by the Biophysical Society 0006-3495/98/08/948/09 $2.00
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