A 1H NMR determination of the solution conformation of a synthetic peptide analogue of calcium-binding site III of rabbit skeletal troponin C.
NMR techniques have been used to determine the structure in solution of acetyl (Asp 105) skeletal troponin C (103-115) amide, one of a series of synthetic peptide analogues of calcium-binding site III of rabbit skeletal troponin C [Marsden et al. (1988) Biochemistry 27, 4198-4206]. The NMR measurements include 1H-1H nuclear Overhauser enhancements and gadolinium-induced 1H relaxation measurements. The former yield short-range internuclear distances (less than 4 A); the latter, once properly corrected for chemical exchange, yield longer range metal to proton distances (5-10 A). These measurements were then used as pseudo potential energy restraints in energy minimization and molecular dynamics calculations to determine the solution structure. Further information was provided by NMR coupling constants, amide proton exchange rates, and the temperature dependences of amide proton chemical shifts. The solution structure of the peptide analogue is very similar to that of the calcium-binding loop in the protein, the root-mean-square deviation between the backbone atoms being approximately 1.1 A.
Marsden
B J
BJ
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Hodges
R S
RS
Sykes
B D
BD
D000818:Animals; D001665:Binding Sites; D002135:Calcium-Binding Proteins; D001330:Electronic Data Processing; D009682:Magnetic Resonance Spectroscopy; D009132:Muscles; D011487:Protein Conformation; D011817:Rabbits; D012996:Solutions; D013816:Thermodynamics; D014336:Troponin; D019209:Troponin C; D014961:X-Ray Diffraction
D002135:Calcium-Binding Proteins; D012996:Solutions; D014336:Troponin; D019209:Troponin C