title: Purification and characterization of a potent homodimeric guanine-specific ribonuclease from fresh mushroom (Pleurotus tuber-regium) sclerotia.
abstract: From the fresh sclerotia of the mushroom Pleurotus tuber-regium, a potent homodimeric ribonuclease exhibiting a molecular weight of 29 kDa in FPLC-gel filtration and sodium dodecyl sulfate-polyacrylamide gel electrophoresis was isolated. The protein was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel, CM-cellulose and Mono S. It manifested strong ribonucleolytic activity toward Poly G, slight activity toward Poly U and Poly A, and minimal activity toward Poly C. Its optimal pH was determined to be 6.5 when yeast transfer RNA was used as substrate. Its ribonucleolytic activity was resistant to heating at 100 degrees C for 30 min but was inhibited by a number of salts. The protein inhibited cell-free translation in a rabbit reticulocyte lysate with an IC50 of 0.09 nM. Three out of the four amino acid residues at the active site (positions 38-41) of P. ostreatus ribonuclease, YNNF, were also found at positions 17-20 in the P. tuber-regum RNase. However, unlike P. ostreatus RNase, no cysteine residues were detected in the N-terminal sequence.
authors.affiliation: Department of Microbiology, College of Biological Science, China Agricultural University, Beijing, China.
authors.forename: H X
authors.initials: HX
authors.lastname: Wang
authors.forename: T B
authors.initials: TB
authors.lastname: Ng
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D020076:Pleurotus / Q000201:enzymology*; D014176:Protein Biosynthesis / Q000187:drug effects*; D011817:Rabbits; D012260:Ribonucleases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D016415:Sequence Alignment
chemical_list: C436527:Pleurotus tuber-regium ribonuclease; D012260:Ribonucleases