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Modification of crossbridge states by ethylene glycol in insect flight muscle

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Summary

Substitution of ethylene glycol for part of the solvent water changes the mechanical properties, structure and nucleotide binding of glycerol-extracted flight muscle fibres from the waterbugLethocerus. On addition of ethylene glycol the rigor tension falls, rapidly and reversibly. With increasing glycol concentration the effect saturates at a non-zero tension. The isotonic stiffness is unchanged on adding ethylene glycol. Adding MgAMPPNP (adenylylimidodiphosphate) to a muscle fibre in 50% ethylene glycol causes a further rapid tension fall; above 100 µm AMPPNP the tension reaches zero. The isotonic stiffness of restretched muscle is then close to that of a relaxed fibre. Removal of MgAMPPNP from the bathing medium has no immediate mechanical effect. After several hours the isotonic stiffness rises to some extent; on removal of the glycol both tension and stiffness rise to rigor values within one minute.3H-Labelled AMPPNP binds to muscle fibres in 50% ethylene glycol in a similar amount to the number of myosin heads present. The binding is tighter than that in aqueous solution and the nucleotide is only released very slowly. Upon removal of the ethylene glycol nucleotide is rapidly released. X-ray diffraction of muscle in 50% ethylene glycol reveals a highly ordered structure, in which both the 14 nm and the 38 nm layer lines are sharply sampled and are of intermediate values between rigor and relaxation. The two inner equatorial peaks are also of intermediate values. On adding MgAMPPNP the pattern resembles that of relaxed muscle. Upon removal of the nucleotide the pattern does not revert towards rigor but on removal of glycol it does. These results are interpreted in terms of changes within the myosin heads and their array within the filament lattice.

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Author notes

  1. M. L. Clarke

    Present address: MRC Cell Biophysics Unit, 26-29 Drury Lane, WC2B 5RL, London, UK

  2. C. D. Rodger

    Present address: Biology Department, Roehampton Institute of Higher Education, SW15, London, UK

Authors and Affiliations

  1. Biophysics Unit, ARC Institute of Animal Physiology, Babraham, CB2 4AT, Cambridge, UK

    M. L. Clarke, C. D. Rodger & R. T. Tregear

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  1. M. L. Clarke
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  2. C. D. Rodger
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  3. R. T. Tregear
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Clarke, M.L., Rodger, C.D. & Tregear, R.T. Modification of crossbridge states by ethylene glycol in insect flight muscle. J Muscle Res Cell Motil 5, 81–96 (1984). https://doi.org/10.1007/BF00713153

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  • DOI: https://doi.org/10.1007/BF00713153

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