Nuclear magnetic resonance studies on calmodulin: spectral assignments in the calcium-free state.
The 400-MHz proton magnetic resonance spectra of calcium-free scallop testis calmodulin (CaM) and pig brain CaM were observed. Detailed spectral assignments were made by resolution enhancement, spin decoupling, and nuclear Overhauser enhancement (NOE) experiments as well as pH titration. Comparison between spectra of scallop testis CaM and pig brain CaM were also utilized for the assignment. Previous assignments for tyrosine-99, histidine-107, epsilon-trimethyllysine-115, and tyrosine-138 [Seamon, K. B. (1980) Biochemistry 19, 207; Krebs, J., & Carafoli, E. (1982) Eur. J. Biochem. 124, 619] were confirmed. Phenylalanine-99 and threonine-143 of scallop testis CaM were identified. Sixteen methyl resonances from one isoleucine, two valines, nine methionines, and the amino-terminal acetyl group were identified. First-stage assignments were made of resonances arising from seven phenylalanines. The uniquely high field shifted phenylalanine resonance previously reported by Seamon was found to consist of two doublets from the two pairs of delta protons of two phenylalanines. The NOE experiments showed that the two phenylalanines are located closely to each other. The large high-field shifts of these phenylalanines were accounted for the ring-current effects due to their proximity. An isoleucine and a valine of which methyl resonances appear at high fields were found to be situated closely to each other. It was found that two delta protons and two epsilon protons of almost all aromatic residues are magnetically equivalent, suggesting that the local structure of aromatic residues is so flexible as to permit the rapid flipping motion of the ring.
D000818:Animals; D001923:Brain Chemistry; D002118:Calcium; D002135:Calcium-Binding Proteins; D002147:Calmodulin; D002417:Cattle; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D008974:Mollusca; D011485:Protein Binding; D011487:Protein Conformation; D013552:Swine; D013737:Testis
D002135:Calcium-Binding Proteins; D002147:Calmodulin; D002118:Calcium