Interaction of troponin subunits. The interaction between the inhibitory and tropomyosin-binding subunits.

The interaction of troponin-I and troponin-T was demonstrated by circular dichroism and gel filtration. Troponin-I gives a negative circular dichroism band between 300 and 260 nm while troponin-T gives two weak positive bands, one at 290 nm and the other at 263 nm. When troponin-I and troponin-T were mixed, the complex produced a strong negative circular dichroism band with a maximum around 280 nm. This band was most intense with a molar ratio of troponin-T to troponin-I of 1:1. The intensity of the band was 2.4 times that expected from the separate components. The interaction was independent of salt concentration from 0.15 to 0.5 M KCl. Gel filtration on Sephacryl S-200 showed that a stable 1:1 complex was formed between troponin-T and troponin-I. When troponin-C was added to the complex of troponin-T.troponin-I; the reconstituted troponin had a circular dichroism spectrum identical to that of native troponin. The oxidation state of troponin-I was important in reconstituting troponin. Oxidized troponin-I produced less change in the near ultraviolet circular dichroism when added to troponin-T and the troponin-C than did reduced troponin-I. This showed the subunits were not assembled correctly with oxidized troponin-I. When the reconstituted complex was reduced, the circular dichroism was restored to that of native troponin. Troponin reconstituted with oxidized troponin-I did not confer calcium sensitivity on actomyosin ATPase; activity was restored by reducing the complex.