The effect of polyethylene glycol on the mechanics and ATPase activity of active muscle fibers.
We have used polyethylene glycol (PEG) to perturb the actomyosin interaction in active skinned muscle fibers. PEG is known to potentiate protein-protein interactions, including the binding of myosin to actin. The addition of 5% w/v PEG (MW 300 or 4000) to active fibers increased fiber tension and decreased shortening velocity and ATPase activity, all by 25-40%. Variation in [ADP] or [ATP] showed that the addition of PEG had little effect on the dissociation of the cross-bridge at the end of the power stroke. Myosin complexed with ADP and the phosphate analog V(i) or AlF(4) binds weakly to actin and is an analog of a pre-power-stroke state. PEG substantially enhances binding of these states both in active fibers and in solution. Titration of force with increasing [P(i)] showed that PEG increased the free energy available to drive the power stroke by about the same amount as it increased the free energy available from the formation of the actomyosin bond. Thus PEG potentiates the binding of myosin to actin in active fibers, and it provides a method for enhancing populations of some states for structural or mechanical studies, particularly those of the normally weakly bound transient states that precede the power stroke.
Chinn
M K
MK
Department of Biochemistry and Biophysics, Cardiovascular Research Institute, University of California, San Francisco 94143 USA.
Myburgh
K H
KH
Pham
T
T
Franks-Skiba
K
K
Cooke
R
R
D000199:Actins / Q000737:chemistry; D000205:Actomyosin / Q000737:chemistry; D000244:Adenosine Diphosphate / Q000031:analogs & derivatives / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D017607:Aluminum Compounds / Q000494:pharmacology; D000818:Animals; D001696:Biomechanical Phenomena; D005459:Fluorides / Q000494:pharmacology; D007537:Isometric Contraction / Q000187:drug effects; D007700:Kinetics; D018485:Muscle Fibers, Skeletal / Q000187:drug effects*; D009218:Myosins / Q000737:chemistry; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011485:Protein Binding; D016658:Psoas Muscles / Q000187:drug effects*; D011817:Rabbits
D000199:Actins; D017607:Aluminum Compounds; C050992:tetrafluoroaluminate; D011092:Polyethylene Glycols; D000244:Adenosine Diphosphate; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D009218:Myosins; D005459:Fluorides