Monoclonal antibodies detect and stabilize conformational states of smooth muscle myosin.
Antibodies with epitopes near the heavy meromyosin/light meromyosin junction distinguish the folded from the extended conformational states of smooth muscle myosin. Antibody 10S.1 has 100-fold higher avidity for folded than for extended myosin, while antibody S2.2 binds preferentially to the extended state. The properties of these antibodies provide direct evidence that the conformation of the rod is different in the folded than the extended monomeric state, and suggest that this perturbation may extend into the subfragment 2 region of the rod. Two antihead antibodies with epitopes on the heavy chain map at or near the head/rod junction. Magnesium greatly enhances the binding of these antibodies to myosin, showing that the conformation of the heavy chain in the neck region changes upon divalent cation binding to the regulatory light chain. Myosin assembly is also altered by antibody binding. Antibodies that bind to the central region of the rod block disassembly of filaments upon MgATP addition. Antibodies with epitopes near the COOH terminus of the rod, in contrast, promote filament depolymerization, suggesting that this region of the tail is important for assembly. The monoclonal antibodies described here are therefore useful both for detecting and altering conformational states of smooth muscle myosin.
Trybus
K M
KM
Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, MA 02254-9110.
Henry
L
L
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D005895:Gizzard, Avian / Q000378:metabolism; D015151:Immunoblotting; D007700:Kinetics; D051379:Mice; D008815:Mice, Inbred Strains / Q000276:immunology; D008854:Microscopy, Electron; D009130:Muscle, Smooth / Q000378:metabolism*; D009218:Myosins / Q000032:analysis / Q000378:metabolism* / Q000648:ultrastructure; D011487:Protein Conformation; D014422:Turkeys
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000939:Epitopes; D009218:Myosins