Sequential 1H-NMR assignment and solution structure of bovine pancreatic ribonuclease A.
Assignments for 1H-NMR resonances of most of the residues of bovine pancreatic ribonuclease (RNase A) have been obtained by sequence-specific methods. Identification and classification of spin systems have been carried out by two-dimensional phase-sensitive correlated spectroscopy (360 MHz) and single relayed coherence transfer spectroscopy. Sequence-specific assignments have been achieved by phase-sensitive two-dimensional nuclear Overhauser effect spectroscopy. To overcome the problem of spectral overlap use has been made of (a) an exhaustive analysis of partly exchanged RNase A (spectra in D2O), (b) a comparison with the subtilisin-modified enzyme (RNase S) and (c) small spectral perturbations caused by changes in pH and temperature. The secondary structure elements have been identified from the observed sequential, medium and long-range nuclear Overhauser effects together with data from amide-exchange rates. All information collected leads to the conclusion that the crystal and the solution structures are closely similar.
Rico
M
M
Instituto de Estructura de la Materia, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Bruix
M
M
Santoro
J
J
Gonzalez
C
C
Neira
J L
JL
Nieto
J L
JL
Herranz
J
J
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D006859:Hydrogen; D046911:Macromolecular Substances; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008961:Models, Structural; D008969:Molecular Sequence Data; D011487:Protein Conformation; D012259:Ribonuclease, Pancreatic
D046911:Macromolecular Substances; D006859:Hydrogen; D012259:Ribonuclease, Pancreatic