A folded (10 S) conformer of myosin from a striated muscle and its implications for regulation of ATPase activity.
Myosin from the striated adductor muscle of the scallop Pecten maximus is shown to fold into a compact 10 S conformer under relaxing conditions, as has been characterized for smooth and non-muscle myosins. The folding transition is accompanied by the trapping of nucleotide at the active site to give a species with a half-life of about an hour at 20 degrees C. Ca2+ binding to the specific, regulatory sites on a myosin head promotes unfolding to the extended 6 S conformer and activates product release by 60-fold. The unfolding transition, however, remains much slower than the contraction-relaxation cycle of scallop striated muscle and could not play a role in the regulation of these events. The dissociation of products from myosin heads in native thick filaments is Ca2(+)-regulated, but under relaxing conditions the nucleotide is released at least an order of magnitude faster than from the 10 S monomeric myosin, at a rate similar to that observed with heavy meromyosin. Thus, there is no evidence for any intermolecular interaction between neighbouring molecules in the filament analogous to the head-neck intramolecular interaction in the 10 S conformer. It is possible that the 10 S myosin state represents an inert form involved in the control of filament assembly during muscle growth and development. Removal of regulatory light chains or labelling the reactive heavy chain thiol of myosin prevents, or at least disfavours, formation of the folded 10 S conformer and allows separation of the modified protein from the native molecules.
Ankrett
R J
RJ
Department of Biochemistry, University of Leicester, U.K.
Rowe
A J
AJ
Cross
R A
RA
Kendrick-Jones
J
J
Bagshaw
C R
CR
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002850:Chromatography, Gel; D005573:Formycins / Q000378:metabolism; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008974:Mollusca; D009132:Muscles / Q000201:enzymology*; D009218:Myosins / Q000737:chemistry / Q000648:ultrastructure*; D011487:Protein Conformation; D012265:Ribonucleotides / Q000378:metabolism; D012995:Solubility; D013329:Structure-Activity Relationship
D005573:Formycins; D046911:Macromolecular Substances; D012265:Ribonucleotides; C032552:formycin triphosphate; D000255:Adenosine Triphosphate; D009218:Myosins; D002118:Calcium