title: Interaction studies of the 165 000 dalton protein component of the M-line with the S2 subfragment of myosin.
abstract: The M-line protein component of molecular weight 165 000 was isolated and purified from rabbit skeletal muscle using ion exchange chromatography. Sodium dodecyl sulphate electrophoresis revealed that the protein was homogeneous. Circular dichroism measurements indicated that the protein interacts with myosin and heavy meromyosin subfragment 2 (S2). There was an increase in negative ellipticity at 221 nm upon interaction, relative to the calculated values assuming no interprotein interaction. The net increaes in negative ellipticity at 221 nm as a result of interaction of M-protein with myosin and subfragment 2 were 600 degrees and 800 degrees respectively. When the protein was mixed with subfragment 2 in a 1 : 1 mol ratio in 0.5 M KCl/25 mM Tris buffer at pH 8.0, low speed sedimentation equilibrium studies gave a molecular weight of 235 000 +/- 10 000 for the complex, indicative of an interaction of the two components. On a Bio gel A 0.5 m column, M-protein and S2 when applied in 1 : 1 mol ratio, were eluted as a single symmetrical peak and a molecular weight of 230 000 was obtained for the complex from the observed elution volume. Both circular dichroism and sedimentation equilibrium studies indicated no interaction of M-line protein with light meromyosin and subfragment 1. Interaction of the 165 000 component with the flexible hinge region of myosin may have special significance in terms of the mechanism accounting for the reversible expansion of the interfilament distance which occurs during contraction.
mesh_terms: D000818:Animals; D001665:Binding Sites; D002942:Circular Dichroism; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification; D009218:Myosins; D010446:Peptide Fragments; D011485:Protein Binding; D011487:Protein Conformation; D011817:Rabbits; D013056:Spectrophotometry, Ultraviolet
chemical_list: D009124:Muscle Proteins; D010446:Peptide Fragments; D009218:Myosins