Tropomyosin ends determine the stability and functionality of overlap and troponin T complexes.
Tropomyosin binds end to end along the actin filament. Tropomyosin ends, and the complex they form, are required for actin binding, cooperative regulation of actin filaments by myosin, and binding to the regulatory protein, troponin T. The aim of the work was to understand the isoform and structural specificity of the end-to-end association of tropomyosin. The ability of N-terminal and C-terminal model peptides with sequences of alternate alpha-tropomyosin isoforms, and a troponin T fragment that binds to the tropomyosin overlap, to form complexes was analyzed using circular dichroism spectroscopy. Analysis of N-terminal extensions (N-acetylation, Gly, AlaSer) showed that to form an overlap complex between the N-terminus and the C-terminus requires that the N-terminus be able to form a coiled coil. Formation of a ternary complex with the troponin T fragment, however, effectively takes place only when the overlap complex sequences are those found in striated muscle tropomyosins. Striated muscle tropomyosins with N-terminal modifications formed ternary complexes with troponin T that varied in affinity in the order: N-acetylated > Gly > AlaSer > unacetylated. The circular dichroism results were corroborated by native gel electrophoresis, and the ability of the troponin T fragment to promote binding of full-length tropomyosins to filamentous actin.
Palm
Thomas
T
Department of Neuroscience and Cell Biology, University of Medicine and Dentistry of New Jersey-Robert Wood Johnson Medical School, Piscataway, NJ 08854-5635, USA. palmth@umdnj.edu
Greenfield
Norma J
NJ
Hitchcock-DeGregori
Sarah E
SE
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002645:Chickens; D046911:Macromolecular Substances; D020409:Molecular Motor Proteins / Q000737:chemistry*; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D020033:Protein Isoforms / Q000031:analogs & derivatives / Q000737:chemistry; D012680:Sensitivity and Specificity; D013329:Structure-Activity Relationship; D013696:Temperature; D014335:Tropomyosin / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000145:classification; D020107:Troponin T / Q000737:chemistry*
D046911:Macromolecular Substances; D020409:Molecular Motor Proteins; D020033:Protein Isoforms; D014335:Tropomyosin; D020107:Troponin T