Myosin essential light chain isoforms modulate the velocity of shortening propelled by nonphosphorylated cross-bridges.
The differential effects of essential light chain isoforms (LC17a and LC17b) on the mechanical properties of smooth muscle were determined by exchanging recombinant for endogenous LC17 in permeabilized smooth muscle treated with trifluoperazine (TFP). Co-precipitation with endogenous myosin heavy chain verified that 40-60% of endogenous LC17a could be exchanged for recombinant LC17a or LC17b. Upon addition of MgATP in Ca2+-free solution, recombinant LC17 exchange induced slow contractions unaccompanied by regulatory light chain (RLC) phosphorylation only in TFP-treated, but not in untreated, permeabilized smooth muscle; the shortening velocity and rate of force development were approximately 1.5 and 2 times faster, respectively, in response to LC17a than LC17b. Additional incubation with recombinant, thiophosphorylated RLC increased the shortening velocity, independent of the LC17 isoform exchanged. The LC17-induced contractions of TFP-treated muscles were abolished by prior addition of nonphosphorylated RLC. We suggest that LC17 stiffens the lever arm of myosin and, in the absence of regulation by RLC, permits cross-bridge cycling without requiring RLC phosphorylation. Our results are compatible with nonphosphorylated RLC acting as a repressor and with LC17 isoforms modulating the MgADP affinity and, consequently, rate of cooperative cycling of nonphosphorylated cross-bridges.
Matthew
J D
JD
Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, Virginia 22906-0011, USA.
Khromov
A S
AS
Trybus
K M
KM
Somlyo
A P
AP
Somlyo
A V
AV
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives; D000818:Animals; D001696:Biomechanical Phenomena; D002463:Cell Membrane Permeability; D018613:Microscopy, Confocal; D009068:Movement / Q000502:physiology*; D009119:Muscle Contraction / Q000502:physiology*; D009130:Muscle, Smooth / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D018994:Myosin Light Chains / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D010782:Photolysis; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014268:Trifluoperazine / Q000494:pharmacology; D001743:Urinary Bladder / Q000502:physiology / Q000648:ultrastructure
D018994:Myosin Light Chains; D020033:Protein Isoforms; D013438:Sulfhydryl Compounds; D014268:Trifluoperazine; C029273:P(3)-1-(2-nitro)phenylethyladenosine 5'-triphosphate; D000255:Adenosine Triphosphate