Mapping of the Actomyosin Interfaces

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 91; no. 7; pp. 2772 - 2776
• Main Authors: Eldin, Patrick, Le Cunff, Martine, Vosberg, Hans Peter, Mornet, Dominique, Leger, Jean J.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 29.03.1994; National Acad Sciences; National Academy of Sciences
• Subjects: Actins; Actins - metabolism; Actomyosin - genetics; Actomyosin - metabolism; Adenosine Triphosphate - metabolism; Analytical, structural and metabolic biochemistry; Atomic interactions; ATP-Binding Cassette Transporters; Binding sites; Binding, Competitive; Biochemistry; Biochemistry, Molecular Biology; Biological and medical sciences; Carrier Proteins - chemistry; Carrier Proteins - genetics; Contractile proteins; Crosslinking; Crystal structure; Deoxyribonucleic acid; DNA; Elution; Escherichia coli - genetics; Escherichia coli Proteins; Ethenoadenosine Triphosphate - metabolism; Fundamental and applied biological sciences. Psychology; Holoproteins; Humans; Life Sciences; Maltose - metabolism; Maltose-Binding Proteins; Molecular biology; Monosaccharide Transport Proteins; Myocardium - chemistry; Myosins - genetics; Myosins - metabolism; Nucleotides; Peptide Fragments - genetics; Peptide Fragments - metabolism; Protein Conformation; Proteins; Recombinant Fusion Proteins - chemistry; Rectangles; Resins; Structure-Activity Relationship; Myosin ATPase; Enzyme; Contractile protein; Actins; Microfilament; Hydrolases; Affinity; Recombinant DNA; Spatial structure; Structural analysis
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.91.7.2772

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-Base, K., Smith, R., Tomchick, D. R., Benning, M. M., Winkelmann, D. A., Wesenberg, G. \& Holden, H. M. (1993) Science 261, 50-58].