Studies on Ribonucleases in Takadiastase. I

Sato, Kimihide; Egami, Fujio

doi:10.1093/oxfordjournals.jbchem.a126717

Cited by 228 publications

(59 citation statements)

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“…RNase Rh splits the 3',5'-phosphodiester linkage of RNA (through 2',3'-cyclic phosphodiester as an intermediate) without absolute base specificity. Its base specificity estimated from the time-course of release ofnucleotides from RNA is in the order A>G>C>U and is very similar to that of RNase % [3] or RNase M [4]. The enzyme consists of a single polypeptide chain (Air,.…”

Section: Introductionmentioning

confidence: 99%

Crystal and molecular structure of RNase Rh, a new class of microbial ribonuclease from Rhizopus niveus

et al. 1992

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The crystal structure of RNase Rh, a new class of microbial ribonuelease from Rhi'.opus niveus, has been determined at 2.5 A resolution by the multiple isomorphous replacament method. The crystal structure was refined by simulated annealing with molecular dynamics. The current crystallographic R-t'actor is 0,200 in the 10-2.5 A resolution range, The molecular structure which is completely different from the known sh'uctures of RNase A and RNase T~ consists of six e~.heliees and seven//.strands, belonging to the cc+~ type structure, Two histidine and one glutamie acid resid ues which were predicted as rite most probably functional residues by chemical modification studies are Ibund to be clustered, The sterie nature of the active site taken together with the relevant site.directed mutagenesis experiments (lrie ¢t al,) indicates that: (i) the two histidine residues are the general acid and base: and (it) an aspartic acid residue plays a role of recognizing adenine moiety of the suhstrate. . The enzyme consists of a single polypeptide chain (Air,. 24,000) of" 222 amino acid residues, the sequence of which has been determined by Horiuchi et al. [5].As shown in Fig. 1, the locations of the ten halfcystines of RNase gh are almost the same as in gNase T 2 and RNase M, and the sequences around the two histidine and one glutamic acid residues that are suggested to be involved in the active site by Irie [6] are fairly well conserved. The lowest line of Fig. 1 is the sequence of S2-allelic glycoprotein of Nicotiana alata [7]. It is well known that this kind of glycoprotein is involved in self-incompatibility in flowering plants. Although overall homology between Sa-glycoprotein and the family ofRNase Rh, T2 and M is rather poor (about Abbreviations: RNase, ribonuclease; r.m.s,, root.mean.square.

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Section: Introductionmentioning

confidence: 99%

Crystal and molecular structure of RNase Rh, a new class of microbial ribonuclease from Rhizopus niveus

et al. 1992

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“…Hydrolysis of the samples was routinely performed in constantly boiling HCI containing 0.2% phenol at 110°C for 24 h. Low recovery is due to the HC1 hydrolysis. For an explanation of this table see the legend to Table 2s Amino acid For an explanation of this table see the legend to Table 2s Amino acid Ch-1 Ch-2 Ch-3-Ch-3-Ch-4 Ch-5 Ch-6 Ch-7-Ch-7-Ch-8 Ch-9 Ch-10 Ch-11 Ch-12 Ch- (1) 0.9 (1) 0.7 (1)…”

Section: Extraction Of Rnase T2 and Removal Of Rnase T I Andmentioning

confidence: 99%

Amino‐acid sequence of ribonuclease T₂ from Aspergillus oryzae

Kawata

Sakiyama

Tamaoki

1988

European Journal of Biochemistry

130

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The amino acid sequence of ribonuclease T2 (RNase T2) from Aspergillus oryzae has been determined. This has been achieved by analyzing peptides obtained by digestions with Achromobacter lyticus protease I, Staphylococcus aureus V8 protease, and a-chymotrypsin of two large cyanogen bromide peptides derived from the reduced and S-carboxymethylated or S-aminoethylated protein. Digestion with A . lyticus protease I was successfully used to degrade the N-terminal half of the S-aminoethylated protein at cysteine residues.RNase T2 is a glycoprotein consisting of 239 amino acid residues with a relative molecular mass of 29155. The sugar content is 7.9% (by mass). Three glycosylation sites were determined at Asns 1 5 7 6 and 239. Apparently RNase T2 has a very low degree of sequence similarity with RNase T I , but a considerable similarity is observed around the amino acid residues involved in substrate recognition and binding in RNase TI. These similar residues may be important for the catalytic activity of RNase T2.RNase T2 was first reported by Sato and Egami [l] as a ribonuclease in Takadiastase produced from a culture extract of Aspergillus oryzae and purified independently by Rushizky and Sober [2] and by Uchida [3]. The relative molecular mass, M,, of RNase T2 was estimated to be 30500 [2] or 36000 [3] including about 10% sugars. RNase T2 exerts its catalytic activity on the phosphodiester bonds of RNA with a preference for adenylic acid residues without an absolute base specificity [2, 31. In addition to RNase TZ, RNase T I was also purified from Takadiastase. RNase TI, a small protein with an M , 11 000, has a strict substrate specificity for guanylic acid, and Glu58 [4], His40 and His92 [5] have been shown to be essential for catalysis. It is therefore important to examine the similarity and dissimilarity of the structure and function of RNases T2 and TI, which are produced by the same fungus but differ from each other with respect to molecular size and substrate specificity.Although the primary structures of several smaller ribonucleases including RNase TI are known [6], there is no known primary structure of a single-chain ribonuclease with M , = 20000-30000. We have undertaken the elucidation of the primary structure of RNase T2 as the first step toward gaining an insight into the molecular basis of catalysis with a ribonuclease of the above size. In this paper, we describe the complete amino acid sequence of RNase T2 and compare its primary structure with those of other ribonucleases. MATERIALS AND METHODS MaterialsAchromobacter lyticus protease I and ethyleneimine were kindly donated by Dr T. Masaki (Faculty of Agriculture, Ibaraki University). The following materials were purchased from the indicated sources: Staphylococcus aureus V8 protease from Miles Laboratories; a-chymotrypsin from Sigma; 4 M met hanesulfonic acid containing 0.2 % 3-(2-aminoethyl)indole from Pierce; yeast RNA from Tokyo Kasei. All other chemicals were of the highest grade commercially available. Preparation of RNase T2RNase T2 was p...

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“…Acid protease 8 ) and glucoamylase 9 ) from a Monascus fungus have been described. RN ase is produced by organisms in the genera Aspergillus, [10][11][12][13][14][15][16] Rhizopus,17) Neurospora, 18) and Trichoderma. 19) In 1964, Saruno et al 20 ) reported the finding of RNA-degrading enzymes in the genus Monascus but their purification and enzymatic properties were not reported.…”

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confidence: 99%

Purification and Properties of a Ribonuclease from a Species of the GenusMonascus

Yasuda¹,

Ikehara²,

Tawata³

et al. 1995

Bioscience, Biotechnology, and Biochemistry

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Studies on Ribonucleases in Takadiastase. I

Cited by 228 publications

References 1 publication

Crystal and molecular structure of RNase Rh, a new class of microbial ribonuclease from Rhizopus niveus

Crystal and molecular structure of RNase Rh, a new class of microbial ribonuclease from Rhizopus niveus

Amino‐acid sequence of ribonuclease T₂ from Aspergillus oryzae

Purification and Properties of a Ribonuclease from a Species of the GenusMonascus

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Studies on Ribonucleases in Takadiastase. I

Cited by 228 publications

References 1 publication

Crystal and molecular structure of RNase Rh, a new class of microbial ribonuclease from Rhizopus niveus

Crystal and molecular structure of RNase Rh, a new class of microbial ribonuclease from Rhizopus niveus

Amino‐acid sequence of ribonuclease T2 from Aspergillus oryzae

Purification and Properties of a Ribonuclease from a Species of the GenusMonascus

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Amino‐acid sequence of ribonuclease T₂ from Aspergillus oryzae