Filamin and gelsolin influence Ca(2+)-sensitivity of smooth muscle thin filaments.
Sheep aorta thin filaments were prepared by ultracentrifugation of an ATP-containing extract in the presence of different concentrations of ethanediol. Thin filaments prepared without ethanediol contained small quantities of tropomyosin (0.027 Tm:actin) and caldesmon (0.017 CD:actin) and activated the MgATPase of skeletal myosin independently of Ca2+. Ultracentrifugation in the presence of 10-20% ethanediol resulted in preparation of thin filaments with increased content of tropomyosin (0.17 Tm:actin) and caldesmon (0.04 CD:actin). These thin filaments possessed high Ca(2+)-sensitivity in activation of skeletal muscle myosin ATPase. Besides actin, tropomyosin and caldesmon, thin filaments contained gelsolin and filamin. Gelsolin content (0.007 gelsolin:actin) was independent of the presence of ethanediol. The filamin content decreased from 0.015 to 0.007 mol:mol actin when the ethanediol concentration was increased from 0 to 20%, and was negatively correlated with the Ca2+ sensitivity of thin filaments. In a reconstituted system, pure filamin or gelsolin affected caldesmon regulation of actomyosin ATPase. Gelsolin (0.01:actin) reduced the inhibition of actomyosin ATPase caused by caldesmon and increased the potency of Ca(2+)-calmodulin in reversing this inhibition. Filamin (0.007:actin) also decreased the inhibitory action of caldesmon on actin-activated myosin ATPase and also potentiated the reversal of this inhibition by calmodulin. We conclude that minor components of smooth muscle thin filaments (gelsolin and filamin) significantly modify caldesmon mediated regulation of actomyosin ATPase. We suggest a tropomyosin-mediated mechanism by which filamin or gelsolin could exert similar effects.
Gusev
N B
NB
Department of Biochemistry, School of Biology, Moscow State University, Russia.
Pritchard
K
K
Hodgkinson
J L
JL
Marston
S B
SB
D008841:Actin Cytoskeleton / Q000187:drug effects* / Q000502:physiology; D000205:Actomyosin / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D003285:Contractile Proteins / Q000494:pharmacology*; D004789:Enzyme Activation; D064448:Filamins; D018260:Gelsolin / Q000494:pharmacology*; D008840:Microfilament Proteins / Q000494:pharmacology*; D009119:Muscle Contraction / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000502:physiology; D009218:Myosins / Q000378:metabolism*; D012756:Sheep; D014335:Tropomyosin / Q000378:metabolism
D002148:Calmodulin-Binding Proteins; D003285:Contractile Proteins; D064448:Filamins; D018260:Gelsolin; D008840:Microfilament Proteins; D014335:Tropomyosin; D000205:Actomyosin; D009218:Myosins; D002118:Calcium