Mapping of the actomyosin interfaces.
Recombinant DNA methods were used to obtain soluble, undenatured fragments of the heavy chain of myosin subfragment 1 (S-1). These fragments were of preselected lengths and could include protease-sensitive segments that are destroyed when other preparation methods are used. Actin binding by each of the three contiguous segments (residues 1-248, 249-524, and 518-722, essentially spanning the entire S-1 heavy chain) was demonstrated. ATP binding, comparable to that of native S-1, was obtained only with a segment consisting of residues 1-524. Competition among the various fragments for actin was also studied. The data are discussed in relation to the recently reported resolved structure of S-1 [Rayment, I., Rypnieski, R. W., Schmidt-Bäse, K., Smith, R., Tomchick, D. R., Benning, M. M., Winkelmann, D. A., Wesenberg, G. & Holden, H. M. (1993) Science 261, 50-58].
Eldin
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Institut National de la Santé et de la Recherche Médicale, Faculté de Pharmacie Bât. K, Montpellier, France.
Le Cunff
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M
Vosberg
H P
HP
Mornet
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D
Léger
J J
JJ
D018528:ATP-Binding Cassette Transporters; D000199:Actins / Q000378:metabolism*; D000205:Actomyosin / Q000235:genetics / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D001667:Binding, Competitive; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D004985:Ethenoadenosine Triphosphate / Q000378:metabolism; D006801:Humans; D008320:Maltose / Q000378:metabolism; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D009206:Myocardium / Q000737:chemistry; D009218:Myosins / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism*; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship
D018528:ATP-Binding Cassette Transporters; D000199:Actins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D004985:Ethenoadenosine Triphosphate; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli; D008320:Maltose; D000255:Adenosine Triphosphate; D000205:Actomyosin; D009218:Myosins