Cross-bridge binding to actin and force generation in skinned fibers of the rabbit psoas muscle in the presence of antibody fragments against the N-terminus of actin.
To assess the significance of the NH2-terminus of actin for cross-bridge action in muscle, skinned fibers of rabbit psoas muscle were equilibrated with Fab fragments of antibodies directed against the first seven N-terminal residues of actin. With the antibody fragment, active force is more inhibited than relaxed fiber stiffness, or stiffness in rigor or in the presence of magnesium pyrophosphate. Inhibition of stiffness in rigor or with magnesium pyrophosphate does not necessarily indicate involvement of the NH2-terminus of actin in strong cross bridge binding to actin but may simply result from the large size of the Fab. At high Fab concentrations, active force is essentially abolished, whereas stiffness is still detectible under all conditions. Thus, complete inhibition of active force apparently is not due to interference with cross-bridge binding to actin but may result from the Fab-mimicking inhibition of the thin filament by Troponin-1 binding to the NH2-terminus of actin at low Ca2+. However, although Troponin-1 is released from the NH2-terminus at high Ca2+, the Fab is not, thus disallowing force generation upon increase in Ca2+. These data are consistent with involvement of the NH2-terminus of actin in both weak cross-bridge binding to actin and Ca2+ regulation of the thin filament.
Brenner
B
B
Department of Clinical Physiology, Medical School Hannover, Germany.
Kraft
T
T
DasGupta
G
G
Reisler
E
E
D000199:Actins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D002118:Calcium / Q000494:pharmacology; D011756:Diphosphates / Q000494:pharmacology; D007140:Immunoglobulin Fab Fragments; D066298:In Vitro Techniques; D007700:Kinetics; D017616:Magnesium Compounds / Q000494:pharmacology; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D009218:Myosins / Q000378:metabolism; D016658:Psoas Muscles / Q000276:immunology / Q000378:metabolism / Q000502:physiology; D011817:Rabbits; D012518:Sarcomeres / Q000378:metabolism
D000199:Actins; D011756:Diphosphates; D007140:Immunoglobulin Fab Fragments; D017616:Magnesium Compounds; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; C034732:magnesium pyrophosphate; D009218:Myosins; D002118:Calcium