Isolation, properties and primary structure of coypu and chinchilla pancreatic ribonuclease.

Pancreatic ribonucleases from the hystricomorph rodent species: coypu and chinchilla were isolated using chromatography on carboxymethyl-cellulose. The amino acid sequences were determined from tryptic digests of the aminoethylated proteins. The tryptic peptides were positioned in the sequence by homology with other pancreatic ribonucleases. Coypu pancreas contains one carbohydrate-containing ribonuclease component. From chinchilla pancreas two carbohydrate-containing ribonuclease components were obtained; one homogeneous and the other heterogeneous. The latter differs from the first in being more acidic; it exhibits heterogeneity both in its carbohydrate moiety (glycopeptides both with and without sialic acid were isolated) and in amino acid sequence (probably glycine at position 32 has been partially substituted by aspartic acid). In both ribonucleases the carbohydrate is attached to asparagine 34. Earlier results on the titration behaviour of histidine residues in both proteins obtained by nuclear magnetic resonance spectroscopy are discussed. An ion bridge between the invariant glutamic acid 49 and histidine 80 may explain the high pK value of the latter.