Influence of the cardiac myosin hinge region on contractile activity.
The participation of cardiac myosin hinge in contractility was investigated by in vitro motility and ATPase assays and by measurements of sarcomere shortening. The effect on contractile activity was analyzed using an antibody directed against a 20-amino acid peptide within the hinge region of myosin. This antibody bound specifically at the hinge at a distance of 55 nm from the S1/S2 junction, was specific to human, dog, and rat cardiac myosins, did not crossreact with gizzard or skeletal myosin, and had no effect on ATPase activity of purified S1 and myofibrils. However, it completely suppressed the movement of actin filaments in in vitro motility assays and reduced active shortening of sarcomeres of skinned cardiac myocytes by half. Suppression of motion by the anti-hinge antibody may reflect a mechanical constraint imposed by the antibody upon the mobility of the S2 region of myosin. The results suggest that the steps in the mechanochemical energy transduction can be separately influenced through S2.
Margossian
S S
SS
Department of Biochemistry, Montefiore Medical Center, Bronx, NY 10467.
Krueger
J W
JW
Sellers
J R
JR
Cuda
G
G
Caulfield
J B
JB
Norton
P
P
Slayter
H S
HS
D000595:Amino Acid Sequence; D000818:Animals; D000936:Antigen-Antibody Complex; D004285:Dogs; D005895:Gizzard, Avian; D006321:Heart / Q000502:physiology; D006801:Humans; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000502:physiology; D009218:Myosins / Q000235:genetics / Q000276:immunology / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D010455:Peptides / Q000138:chemical synthesis; D011863:Radioimmunoassay; D051381:Rats; D012518:Sarcomeres / Q000502:physiology; D014422:Turkeys
D000936:Antigen-Antibody Complex; D010455:Peptides; D009218:Myosins