The C-terminus of troponin T is essential for maintaining the inactive state of regulated actin.
Striated muscle contraction is regulated by the actin binding proteins tropomyosin and troponin. Defects in these proteins lead to myopathies and cardiomyopathies. Deletion of the 14 C-terminal residues of cardiac troponin T leads to hypertrophic cardiomyopathy. We showed earlier that regulated actin containing Δ14 TnT was more readily activated than wild-type regulated actin. We suggested that the equilibria among the inactive (blocked), intermediate (closed or calcium), and active (open or myosin) states was shifted to the active state. We now show that, in addition, such regulated actin filaments cannot enter the inactive or blocked state. Regulated actin containing Δ14 TnT had ATPase activities in the absence of Ca2+ that were higher than wild-type filaments but far below the fully active rate. The rapid dissociation of S1-ATP from regulated actin filaments containing Δ14 TnT and acrylodan-labeled tropomyosin did not show the fluorescence increase characteristic of moving to the inactive state. Replacing wild-type TnI with S45E TnI, that favors the inactive state, did not restore the fluorescence change. We conclude that TnT has a previously unrecognized role in forming the inactive state of regulated actin.
Department of Biochemistry and Molecular Biology, Brody School of Medicine, East Carolina University, Greenville, North Carolina, USA.
Andrew J
AJ
Franklin
Tamatha
T
Baxley
Tomoyoshi
T
Kobayashi
Joseph M
JM
Chalovich
D015081:2-Naphthylamine / Q000031:analogs & derivatives / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D005453:Fluorescence; D006801:Humans; D007700:Kinetics; D008027:Light; D008954:Models, Biological; D018482:Muscle, Skeletal / Q000201:enzymology; D050505:Mutant Proteins / Q000378:metabolism; D009218:Myosins / Q000378:metabolism; D012542:Scattering, Radiation; D013329:Structure-Activity Relationship; D013997:Time Factors; D014335:Tropomyosin / Q000378:metabolism; D020107:Troponin T / Q000737:chemistry* / Q000378:metabolism*
D000199:Actins; D050505:Mutant Proteins; D014335:Tropomyosin; D020107:Troponin T; C038518:acrylodan; D000255:Adenosine Triphosphate; D015081:2-Naphthylamine; D009218:Myosins; D005033:Ethylmaleimide; D002118:Calcium