The influence of myosin converter and relay domains on cross-bridge kinetics of Drosophila indirect flight muscle.
We are investigating the influence of the converter and relay domains on elementary rate constants of the actomyosin cross-bridge cycle. The converter and relay domains vary between Drosophila myosin heavy chain isoforms due to alternative mRNA splicing. Previously, we found that separate insertions of embryonic myosin isoform (EMB) versions of these domains into the indirect flight muscle (IFM) myosin isoform (IFI) both decreased Drosophila IFM power and slowed muscle kinetics. To determine cross-bridge mechanisms behind the changes, we employed sinusoidal analysis while varying phosphate and MgATP concentrations in skinned Drosophila IFM fibers. Based on a six-state cross-bridge model, the EMB converter decreased myosin rate constants associated with actin attachment and work production, k(4), but increased rates related to cross-bridge detachment and work absorption, k(2). In contrast, the EMB relay domain had little influence on kinetics, because only k(4) decreased. The main alteration was mechanical, in that work production amplitude decreased. That both domains decreased k(4) supports the hypothesis that these domains are critical to lever-arm-mediated force generation. Neither domain significantly influenced MgATP affinity. Our modeling suggests the converter domain is responsible for the difference in rate-limiting cross-bridge steps between EMB and IFI myosin--i.e., a myosin isomerization associated with MgADP release for EMB and Pi release for IFI.
Yang
Chaoxing
C
Department of Biology and Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York, USA.
Kaplan
Charlotte N
CN
Thatcher
Maria L
ML
Swank
Douglas M
DM
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D029721:Drosophila Proteins / Q000737:chemistry* / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism* / Q000502:physiology; D005426:Flight, Animal; D007700:Kinetics; D008958:Models, Molecular; D018482:Muscle, Skeletal / Q000378:metabolism*; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary
D029721:Drosophila Proteins; D010710:Phosphates; D020033:Protein Isoforms; D000255:Adenosine Triphosphate; D009218:Myosins
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