The converter domain modulates kinetic properties of Drosophila myosin.
Recently the converter domain, an integral part of the "mechanical element" common to all molecular motors, was proposed to modulate the kinetic properties of Drosophila chimeric myosin isoforms. Here we investigated the molecular basis of actin filament velocity (V(actin)) changes previously observed with the chimeric EMB-IC and IFI-EC myosin proteins [the embryonic body wall muscle (EMB) and indirect flight muscle isoforms (IFI) with genetic substitution of the IFI and EMB converter domains, respectively]. In the laser trap assay the IFI and IFI-EC myosins generate the same unitary step displacement (IFI = 7.3 +/- 1.0 nm, IFI-EC = 5.8 +/- 0.9 nm; means +/- SE). Thus converter-mediated differences in the kinetics of strong actin-myosin binding, rather than the mechanical capabilities of the protein, must account for the observed V(actin) values. Basal and actin-activated ATPase assays and skinned fiber mechanical experiments definitively support a role for the converter domain in modulating the kinetic properties of the myosin protein. We propose that the converter domain kinetically couples the P(i) and ADP release steps that occur during the cross-bridge cycle.
Littlefield
Kimberly Palmiter
KP
Department of Biology, Molecular Biology Institute, San Diego State University, San Diego, California 92182-4614, USA.
Swank
Douglas M
DM
Sanchez
Becky M
BM
Knowles
Aileen F
AF
Warshaw
David M
DM
Bernstein
Sanford I
SI
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000818:Animals; D004330:Drosophila / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005426:Flight, Animal / Q000502:physiology; D007700:Kinetics; D009132:Muscles / Q000196:embryology / Q000378:metabolism; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D000251:Adenosine Triphosphatases; D009218:Myosins