Modification of the interactions of myosin with actin and 5'-adenylyl imidodiphosphate by substitution of ethylene glycol for water.
We studied the effect of replacing water by ethylene glycol as solvent on the properties of skeletal muscle myosin, myosin subfragment-1 (S1) and heavy meromyosin. Ethylene glycol (50%, v/v) had no detectable effect on the affinity of myosin or actomyosin for the substrate analogue 5'-adenylyl imidodiphosphate (AMPPNP). However, the rate constants for formation and dissociation of the myosin X MgAMPPNP complex were reduced 200-fold; the logarithm of the dissociation rate was roughly proportional to the fractional concentration of ethylene glycol. Nucleotide dissociation was accelerated at least 300-fold by pure actin but remained slow with regulated actin in the absence of Ca2+. Ethylene glycol substitution reduced the affinity of S1 and the S1 X MgAMPPNP complex for actin equally (100-fold at 50% ethylene glycol). These results show that ethylene glycol has specific effects on myosin's enzymic mechanism, which can account for its effect on the tension and stiffness of glycerinated muscle fibres.
D000199:Actins / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000037:antagonists & inhibitors; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000378:metabolism*; D017301:Ca(2+) Mg(2+)-ATPase; D002850:Chromatography, Gel; D019855:Ethylene Glycol; D005026:Ethylene Glycols / Q000494:pharmacology*; D006868:Hydrolysis; D007700:Kinetics; D046911:Macromolecular Substances; D015879:Myosin Subfragments; D009218:Myosins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D012997:Solvents / Q000494:pharmacology*
D000199:Actins; D005026:Ethylene Glycols; D046911:Macromolecular Substances; D015879:Myosin Subfragments; D010446:Peptide Fragments; D012997:Solvents; D000266:Adenylyl Imidodiphosphate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D009218:Myosins; D019855:Ethylene Glycol