Localization of a myosin subfragment-1 interaction site on the C-terminal part of actin.
The actin-myosin head complex in the rigor state reveals several high-affinity sites on the actin molecule in sequences 18-28 and 40-113. In the presence of Mg(2+)-ATP, participation of the actin N-terminal 1-7 sequence is known to occur. The proximity of the C-terminal region of actin to the A1 light chain of the myosin head [S-1(A1)] (where S-1 is myosin subfragment-1) was described previously. We observed that C-terminal antigenic structures located near Met-305, Met-325 and Met-355 and the C-terminal end (Cys-374) of actin are markedly modified in the presence of S-1(A1), S-1(A2) and scallop S-1 and in the absence of Mg(2+)-ATP. This seems to rule out any important specific involvement of the A1 light chain in the described conformational changes. An S-1-binding site was located in this actin C-terminal region by testing the tryptic CB9 peptide (360-372 sequence) previously implicated in the A1 light chain interaction. This peptide was able to bind well to S-1(A1), S-1(A2) and scallop S-1, but not in the presence of Mg(2+)-pyrophosphate. These results strengthen the hypothesis of a multisite interface between S-1 and actin located in the actin subdomain I.
Labbé
J P
JP
Centre de Recherches de Biochimie Macromoléculaire U.P.R. (C.N.R.S.), Unité 249 (I.N.S.E.R.M.), Montpellier, France.
Boyer
M
M
Roustan
C
C
Benyamin
Y
Y
D000199:Actins / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D000906:Antibodies; D001665:Binding Sites; D006801:Humans; D007120:Immunochemistry; D046911:Macromolecular Substances; D015879:Myosin Subfragments / Q000032:analysis / Q000378:metabolism*; D009218:Myosins / Q000032:analysis / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D011487:Protein Conformation; D011817:Rabbits; D014357:Trypsin / Q000378:metabolism
D000199:Actins; D000906:Antibodies; D046911:Macromolecular Substances; D015879:Myosin Subfragments; D010446:Peptide Fragments; D014357:Trypsin; D009218:Myosins