Proton NMR assignments and regular backbone structure of bovine pancreatic ribonuclease A in aqueous solution

• Published in: Biochemistry (Easton) 1989-07, Vol.28 (14), p.5930-5938
• Main Authors: Robertson, Andrew D, Purisima, Enrico O, Eastman, Margaret A, Scheraga, Harold A
• Format: Article
• Language: English
• Subjects: 550601 - Medicine- Unsealed Radionuclides in Diagnostics; Amino Acid Sequence; AMINO ACIDS; Analytical, structural and metabolic biochemistry; ANIMALS; AQUEOUS SOLUTIONS; BARYONS; Biological and medical sciences; BODY; CARBOXYLIC ACIDS; CATTLE; CHEMICAL SHIFT; COHERENT SCATTERING; DIFFRACTION; DIGESTIVE SYSTEM; DISPERSIONS; DOMESTIC ANIMALS; ELEMENTARY PARTICLES; ENDOCRINE GLANDS; ENZYMES; Enzymes and enzyme inhibitors; ESTERASES; FERMIONS; Fundamental and applied biological sciences. Psychology; GLANDS; HADRONS; HYDROLASES; HYDROXY ACIDS; LEUCINE; MAGNETIC RESONANCE; Magnetic Resonance Spectroscopy; MAMMALS; MIXTURES; Molecular Sequence Data; N.M.R; NUCLEAR MAGNETIC RESONANCE; NUCLEONS; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANS; OVERHAUSER EFFECT; PANCREAS; Pancreas - enzymology; PHOSPHODIESTERASES; Protein Conformation; PROTONS; RADIOLOGY AND NUCLEAR MEDICINE; RESONANCE; Ribonuclease, Pancreatic; RNA-ASE; RUMINANTS; SCATTERING; SERINE; SOLUTIONS; THREONINE; VALINE; VERTEBRATES; Water; X-RAY DIFFRACTION
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi00440a033

Proton NMR assignments have been made for 121 of the 124 residues of bovine pancreatic ribonuclease A (RNase A). During the first stage of assignment, COSY and relayed COSY data were used to identify 40 amino acid spin systems belonging to alanine, valine, threonine, isoleucine, and serine residues. Approximately 60 other NH-alpha CH-beta CH systems were also identified but not assigned to specific amino acid type. NOESY data then were used to connect sequentially neighboring spin systems; approximately 475 of the possible 700 resonances in RNase A were assigned in this way. Our assignments agree with those for 20 residues assigned previously [Hahn, U., & Rüterjans, H. (1985) Eur. J. Biochem. 152, 481-491]. Additional NOESY correlations were used to identify regular backbone structure elements in RNase A, which are very similar to those observed in X-ray crystallographic studies [Wlodawer, A., Borkakoti, N., Moss, D. S., & Howlin, B. (1986) Acta Crystallogr. B42, 379-387].