The distance between the high affinity sites of troponin-C measured by interlanthanide ion energy transfer.
Trivalent lanthanide ions are known to be good substituents for Ca2+ at all four calcium-binding sites of rabbit skeletal troponin-C (TnC). In particular, the visible luminescence of terbium ions bound at the two high affinity sites can be excited via a tyrosine residue. We have carried out energy transfer measurements using Tb3+ as the donor and a number of lanthanide ions as acceptors in order to measure the distance between the two high affinity sites in TnC. Luminescence decay measurements showed that, in the absence of acceptors, TnC-bound Tb3+ decays with a single lifetime of 1.31 ms. In the presence of a good acceptor such as Nd3+, the decay was resolvable into two components of roughly equal amplitude. The first was the same in lifetime as that of TnC-bound Tb3+ alone; the second has a shorter lifetime, presumably due to interlanthanide ion energy transfer. From these lifetimes and published critical transfer distances (Horrocks, W. DeW., Jr., Rhee, M.-J., Snyder, A.P., and Sudnick, D.R. (1980) J. Am. Chem. Soc. 102, 3650-3652), we obtained a distance of 0.92 nm between the two high affinity sites. This distance is consistent with the fact that the two high affinity sites have been located in the COOH-terminal half of TnC and with the proposal that each half of TnC is homologous in structure to parvalbumin (Kretsinger, R. H., and Barry, C. D. (1975) Biochim. Biophys. Acta 405, 40-52).
D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D004735:Energy Transfer; D008163:Luminescent Measurements; D008674:Metals, Rare Earth; D009124:Muscle Proteins / Q000378:metabolism*; D009132:Muscles / Q000378:metabolism; D009354:Neodymium; D011485:Protein Binding; D011817:Rabbits; D013725:Terbium; D014336:Troponin / Q000378:metabolism*; D019209:Troponin C
D008674:Metals, Rare Earth; D009124:Muscle Proteins; D014336:Troponin; D019209:Troponin C; D013725:Terbium; D009354:Neodymium; D002118:Calcium