title: The secondary structure of a pyrimidine-guanine sequence-specific ribonuclease possessing cytotoxic activity from the oocytes of Rana catesbeiana.
abstract: RC-RNase is a pyrimidine-guanine sequence-specific ribonuclease and a sialic-acid-binding lectin purified from Rana catesbeiana (bullfrog) oocytes. This 111-amino acid protein exhibits cytotoxicity toward several tumor cell lines. In this paper we report the assignments of proton NMR resonances and the identification of the secondary structure deduced from NOE constraints, chemical shift index, 3JNH alpha and amide proton exchange rates. The protein was directly isolated from bullfrog oocytes; we were able to assign all but five of the amino acid backbone protons of the unlabeled protein by analyzing a large set of two-dimensional proton NMR spectra obtained at several temperatures and pH conditions. Our results indicate that the structure of RC-RNase is dominated by the presence of two triple-stranded antiparallel beta-sheets and three alpha-helices, similar to those of the pyrimidine family ribonucleases. Two sets of resonances were observed for 11 amide protons and 8 alpha-protons located in the loop-1 region, an alpha 2 helix, and three beta-strands, (beta 1, beta 3 and beta 4), suggesting the presence of nonlocalized multiple conformations for RC-RNase.
authors.affiliation: Division of Structural Biology, Academia Sinica, Nankang, Taipei, Taiwan, Republic of China.
authors.forename: C
authors.initials: C
authors.lastname: Chen
authors.forename: K
authors.initials: K
authors.lastname: Hom
authors.forename: R F
authors.initials: RF
authors.lastname: Huang
authors.forename: P J
authors.initials: PJ
authors.lastname: Chou
authors.forename: Y D
authors.initials: YD
authors.lastname: Liao
authors.forename: T
authors.initials: T
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D005260:Female; D006147:Guanine; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009865:Oocytes / Q000201:enzymology*; D017433:Protein Structure, Secondary; D011743:Pyrimidines; D011892:Rana catesbeiana; D012260:Ribonucleases / Q000737:chemistry*; D016415:Sequence Alignment; D017421:Sequence Analysis
chemical_list: D011743:Pyrimidines; D006147:Guanine; D012260:Ribonucleases; C030986:pyrimidine