Effect of actin on the tryptic digestion of myosin subfragment 1 in the weakly attached state.
The structure of myosin subfragment 1 (S1) in the weakly attached complex with actin was studied at three specific sites, at the 50-kDa/20-kDa and 27-kDa/50-kDa junctions, and at the N-terminal region, using tryptic digestion as a structure-exploring tool. The structure of S1 at the vicinity of the 50-kDa/20-kDa junction is pH dependent in the weakly attached state because the tryptic cleavage at this site was fully protected by actin at pH 6.2, but the protection was only partial at pH 8.0. Since the actin protection is complete in rigor at both pH values, the results indicate that the structure of S1 at the 50-kDa/20-kDa junction differs in the two states at pH 8.0, but not at pH 6.2. Actin restores the ADP-suppressed tryptic cleavage after Lys213 at the 27-kDa/50-kDa junction in the strongly attached state, but not in the weakly attached state, which indicates structural difference between the two states at this site. ATP and ADP open a new site for tryptic cleavage in the N-terminal region of the S1 heavy chain between Arg23 and Ile24. Actin was found to suppress this cleavage in both weakly and strongly attached states, which shows that, in the vicinity of this site, the structure of S1 is similar in both states. The results indicate that the binding of S1 to actin induces localized changes in the S1 structure, and the extent of these changes is different in the various actin-S1 complexes.
Blotnick
E
E
Department of Oral Biology, Hebrew University, Hadassah School of Dental Medicine, Jerusalem, Israel.
Muhlrad
A
A
D000199:Actins / Q000302:isolation & purification / Q000494:pharmacology*; D007700:Kinetics; D008970:Molecular Weight; D015879:Myosin Subfragments / Q000378:metabolism*; D009218:Myosins / Q000302:isolation & purification / Q000378:metabolism; D010446:Peptide Fragments / Q000302:isolation & purification; D011485:Protein Binding; D014357:Trypsin / Q000378:metabolism*
D000199:Actins; D015879:Myosin Subfragments; D010446:Peptide Fragments; D014357:Trypsin; D009218:Myosins