Electron microscopic mappings of myosin head with site-directed antibodies.
Site-directed antibodies were raised against three synthetic peptides whose sequences correspond to a region around the reactive lysine residue and two protease-sensitive regions of subfragment 1 (S1) of skeletal muscle myosin (one at the junction of the 23,000 Mr and 50,000 Mr segments, the J1 junction; and the other at the junction of the 50,000 Mr and 20,000 Mr segments of the heavy chain, the J2 junction). The antisera cross-reacted with intact myosin with titres of 5 x 10(4) (anti-J1 antiserum) and 10(4) (anti-J2 and anti-reactive lysine residue antisera). Site-specific antibodies purified by S1-Sepharose readily bound to myosin. Electron microscopic examinations of antibody-myosin complexes revealed that the J1 and J2 junctions are located 15 nm and 16 nm from the head-rod junction, respectively, while the reactive lysine residue region is 13 nm from the junction.
Sutoh
K
K
Department of Biophysics and Biochemistry, Faculty of Science, University of Tokyo, Japan.
Tokunaga
M
M
Wakabayashi
T
T
D000818:Animals; D000906:Antibodies; D001665:Binding Sites; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008961:Models, Structural; D015879:Myosin Subfragments; D009218:Myosins; D010446:Peptide Fragments
D000906:Antibodies; D007074:Immunoglobulin G; D046911:Macromolecular Substances; D015879:Myosin Subfragments; D010446:Peptide Fragments; D009218:Myosins