Characterization of two forms of base non-specific and adenylic acid preferential ribonuclease from Aspergillus saitoi.
Two forms of RNases (RNase ML and RNase MM) from Aspergillus saitoi which are base non-specific and adenylic acid preferential were separated from each other by DEAE-cellulose column chromatography. They are indistinguishable with respect to enzymatic properties such as base preferability, pH optimum, kinetic constants measured with 2',3'-cUMP and 2',3'-cCMP as substrates, and effects of ionic strength, physical properties such as heat stability, isoelectric point and circular dichroism spectra, amino acid composition and immunological property. They only differ in carbohydrate content. The apparent molecular weight determined by SDS-disc electrophoresis was 36,000 for RNase ML and 32,000 for RNase MM. Both RNases were reduced and carboxymethylated, and then digested with trypsin, separately. Glycopeptides were isolated from the both digests by gelfiltration and paper chromatography. The amino acid compositions of glycopeptides obtained from RNase ML (ML TS-IIC) and that obtained from RNase MM (MM TS-IIIC) were the same. The amino acid sequences of both glycopeptides determined by Edman degradation and carboxypeptidase digestion were also the same. The results indicated that RNase ML and RNase MM were the same protein having different sizes of carbohydrate chains at one site on the molecule.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001230:Aspergillus / Q000201:enzymology*; D002942:Circular Dichroism; D004355:Drug Stability; D004722:Endoribonucleases / Q000302:isolation & purification* / Q000378:metabolism; D006358:Hot Temperature; D005779:Immunodiffusion; D007700:Kinetics; D008970:Molecular Weight; D011487:Protein Conformation
D000596:Amino Acids; D004722:Endoribonucleases; C022538:ribonuclease M