title: Amino acid sequence and characterization of a rice bran ribonuclease.
abstract: A base-nonspecific and acid ribonuclease (RNase Os) belonging to the RNase T2 family was purified from rice bran to a homogeneous state by SDS-PAGE. The primary structure of RNase Os was determined by protein chemistry and molecular cloning. The RNase Os was a simple protein and consisted of 205 amino acid residues. Its molecular weight was 22578 and its amino acid sequence showed that it was most similar to barley RNase among the known RNase T2 family enzymes having 157 amino acid residues identical with barley RNase. However, its N-terminus was blocked by a gamma-pyroglutamyl residue. The optimal pH of RNase Os was around 5.5. The base preference at the B1 and B2 site of RNase Os was estimated from the rates of hydrolysis of 16 dinucleoside phosphates, to be guanine as the case of RNase LE from tomato. RNase Os was successfully expressed from yeast cells using the E. coli yeast expression vector pYE-RNAP.
authors.affiliation: Department of Microbiology, Hoshi College of Pharmacy, Tokyo, Japan. iwama@hoshi.ac.jp
authors.forename: M
authors.initials: M
authors.lastname: Iwama
authors.forename: Y
authors.initials: Y
authors.lastname: Ogawa
authors.forename: M
authors.initials: M
authors.lastname: Yamagishi
authors.forename: T
authors.initials: T
authors.lastname: Itagaki
authors.forename: N
authors.initials: N
authors.lastname: Inokuchi
authors.forename: T
authors.initials: T
authors.lastname: Koyama
authors.forename: R
authors.initials: R
authors.lastname: Imai
authors.forename: K
authors.initials: K
authors.lastname: Ohgi
authors.forename: T
authors.initials: T
authors.lastname: Tsuji
authors.forename: M
authors.initials: M
authors.lastname: Irie
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D018744:DNA, Plant / Q000032:analysis / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology*; D010447:Peptide Hydrolases; D010940:Plant Proteins / Q000378:metabolism; D012260:Ribonucleases / Q000032:analysis*
chemical_list: D018744:DNA, Plant; D007202:Indicators and Reagents; D010940:Plant Proteins; D012260:Ribonucleases; D010447:Peptide Hydrolases