Purification, some properties, and primary structure of a base non-specific ribonuclease from oyster (Crussdstrea grigus).
A ribonuclease (RNase Oy) was purified to homogeneity on SDS-PAGE from the homogenate of oyster (Crussdstrea grigus). The apparent molecular weight estimated from SDS-PAGE was ca. 28 kDa. The pH optimum of the RNase was 5.0. The RNase released mononucleotides from RNA in the order of 3'-GMP, 3'-AMP, and 3'-UMP. The complete amino acid sequence of RNase Oy was determined, mostly by analyzing the peptides generated by BrCN cleavage or digestion by lysylendopeptidase, staphylococcal V8 protease, and alpha-chymotrypsin. The molecular weight of the protein moiety of RNase Oy deduced from the sequence was 24,359. The sequence of RNase Oy contained two typical histidine residues in segments common to the active site of RNase T2 family enzymes. The locations of six half cystine residues among eight were almost superimposable on those of four known plant RNases of RNase T2 family. The sequence homology between RNase Oy and five fungal and four plant RNases amount, to 43-56 amino acid residues. The amino acid sequence of the N-terminal part of RNase Oy is more similar to those of plant RNases than to those of fungal RNases. This RNase is the first RNase T2 family RNase from mollusc whose primary structure has been elucidated.
Watanabe
H
H
Department of Microbiology, Hoshi College of Pharmacy, Tokyo.
Narumi
H
H
Inaba
T
T
Ohgi
K
K
Irie
M
M
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001482:Base Composition; D005591:Chemical Fractionation; D003488:Cyanogen Bromide; D010450:Endopeptidases; D006868:Hydrolysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010125:Ostreidae / Q000737:chemistry / Q000201:enzymology*; D010446:Peptide Fragments / Q000302:isolation & purification; D012260:Ribonucleases / Q000737:chemistry / Q000302:isolation & purification*; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases
D000596:Amino Acids; D010446:Peptide Fragments; D012260:Ribonucleases; D010450:Endopeptidases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D003488:Cyanogen Bromide