Stiffness of carbodiimide-crosslinked glycerinated muscle fibres in rigor and relaxing solutions at high salt concentrations.
In this article, we have applied a crosslinking technique with a water-soluble carbodiimide to single glycerol-extracted muscle fibres from the rabbit. We have measured the stiffness of the fibres in a relaxing solution at high salt concentration. These fibres were crosslinked to varying extents in the rigor state. The relaxing solution caused uncrosslinked crossbridge heads (S1) to detach. High salt concentrations were used because the fibres were not activated by the crosslinked crossbridges under these conditions, although they were at physiological ionic strength. We found a linear correlation between the extent of S1 crosslinking to thin filaments and the stiffness and that the stiffness in the relaxing solution of muscle fibres with all the S1 heads crosslinked to thin filaments was the same as the rigor stiffness of the fibres before crosslinking. We conclude that the sarcomere compliance is mostly a property of the crossbridges (with more than 65% of the crossbridge compliance in the S1 portions and less than 35% in the S2 portion) and little of other sarcomere structures. In an earlier paper [Kimura & Tawada, Biophys. J. 45, 603-10 (1984)], we demonstrated that the S2 portion of the crossbridge was stiff. It then follows that the crossbridge compliance, and thus the sarcomere compliance, is a property of the S1 heads. Assuming that the S1 portion of the crossbridges in rigor strained muscle fibres is bent, we calculated the Young's modulus of the S1 portion and found that it is about 10(2) MN m-2. Because this order of magnitude is reasonable in terms of globular protein elasticity, bending is likely to be the nature of the S1 compliance in rigor muscle fibres.
D000199:Actins / Q000378:metabolism; D000818:Animals; D002234:Carbodiimides / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000494:pharmacology; D005022:Ethyldimethylaminopropyl Carbodiimide / Q000031:analogs & derivatives / Q000494:pharmacology*; D006982:Hypertonic Solutions / Q000494:pharmacology*; D066298:In Vitro Techniques; D008433:Mathematics; D009126:Muscle Relaxation / Q000187:drug effects; D009132:Muscles / Q000187:drug effects / Q000502:physiology*; D009218:Myosins / Q000378:metabolism; D011817:Rabbits; D013997:Time Factors
D000199:Actins; D002234:Carbodiimides; D003432:Cross-Linking Reagents; D006982:Hypertonic Solutions; C041829:1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide methiodide; D009218:Myosins; D005022:Ethyldimethylaminopropyl Carbodiimide