Proteolysis rates of a myosin heavy chain site with papain. Evidence for a combined LC2-filament-mediated mechanism.
In striated muscle myosin, a proteolysis site at the 25-50 kDa junction, susceptible in the filament and efficiently protected by nucleotides, is similarly protected when myosin is monomeric. Kinetic studies at low ionic strength show a close relationship between LC2 cleavage or degradation rate and cleavage of the 25-50 kDa heavy chain site. The myosin-[(T)-LC2'] species forms normal reconstituted filaments but its 25-50 kDa site susceptibility is closer to that of monomeric myosin, thus becoming practically ionic strength-independent. In this species the absence of the LC2 N-terminal segment induces a significantly greater susceptibility of the papain-sensitive site in LC1. In an LC2-depleted myosin the 25-50 kDa site susceptibility also becomes ionic strength-independent, however, the cleavage rates are then closer to that of filaments. Susceptibility in HMM and S1 is also much less dependent on ionic strength with rates intermediary between those of filament and monomer. These observations show that the maximum susceptibility to papain of the 25-50 kDa site requires both the integrity of the LC2 light chain and the filament structure and furthermore provide evidence that: (i) the LC2 N-terminus interacts specifically with some part of the filament; (ii) this interaction induces a specific transconformation in a region close to the ATPase active site; (iii) there is an interrelationship between LC1 and LC2 light chain N-terminal extremities, at least in the filament structure.
Cardinaud
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D000818:Animals; D066298:In Vitro Techniques; D007700:Kinetics; D008970:Molecular Weight; D015879:Myosin Subfragments / Q000378:metabolism; D009218:Myosins / Q000378:metabolism*; D009711:Nucleotides / Q000378:metabolism; D010206:Papain / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism; D011817:Rabbits; D013329:Structure-Activity Relationship
D015879:Myosin Subfragments; D009711:Nucleotides; D010446:Peptide Fragments; D010206:Papain; D009218:Myosins