The active state of the thin filament is destabilized by an internal deletion in tropomyosin.
The function of three of tropomyosin's sequential quasiequivalent regions was studied by deletion from skeletal muscle alpha-tropomyosin of internal residues 49-167. This deletion mutant tropomyosin spans four instead of the normal seven actins, and most of the tropomyosin region believed to interact with troponin is retained and uninterrupted in the mutant. The mutant tropomyosin was compared with a full-length control molecule that was modified to functionally resemble muscle tropomyosin (Monteiro, P. B., Lataro, R. C., Ferro, J. A., and Reinach, F. C. (1994) J. Biol. Chem. 269, 10461-10466). The tropomyosin deletion suppressed the actin-myosin subfragment 1 MgATPase rate and the in vitro sliding of thin filaments over a heavy meromyosin-coated surface. This inhibition was not reversed by troponin plus Ca2+. Comparable tropomyosin affinities for actin, regardless of the deletion, suggest that the deleted region has little interaction with actin in the absence of other proteins. Similarly, the deletion did not weaken binding of the troponin-tropomyosin complex to actin. Furthermore, Ca2+ had a 2-fold effect on troponin-tropomyosin's affinity for actin, regardless of the deletion. Notably, the deletion greatly weakened tropomyosin binding to myosin subfragment 1-decorated actin, with the full-length tropomyosin having a 100-fold greater affinity. The inhibitory properties resulting from the deletion are attributed to defective stabilization of the myosin-induced active state of the thin filament.
Landis
C A
CA
Departments of Internal Medicine and Biochemistry, The University of Iowa, Iowa City, Iowa 52242, USA.
Bobkova
A
A
Homsher
E
E
Tobacman
L S
LS
D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000648:ultrastructure; D000818:Animals; D002417:Cattle; D017353:Gene Deletion; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000648:ultrastructure; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000648:ultrastructure; D014335:Tropomyosin / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure
D011994:Recombinant Proteins; D014335:Tropomyosin