The Isolation and Partial Characterization of Ribonuclease A from Bison Bison

Overview

Journal Biochem J

Specialty Biochemistry

Date 1973 Nov 1

PMID 4772270

Citations 2

Authors

G R Stewart

K J Stevenson

Affiliations

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Abstract

1. Bison ribonuclease was isolated from pancreas glands of Bison bison by acid extraction, (NH(4))(2)SO(4) fractionation, affinity chromatography on Sepharose-5'-(4-aminophenylphosphoryl)uridine 2',3'-phosphate and ion-exchange chromatography on Bio-Rex-70. 2. The selectivity of the affinity column towards bison ribonuclease in heterogeneous protein solutions was greatly improved by employing piperazine buffers at pH5.3, which decreased non-specific interactions of other proteins. Rapid desorption from the affinity column was obtained with sodium phosphate buffer (pH3). 3. Bison ribonuclease has a total amino acid content very similar to ox ribonuclease. Inactivation of bison ribonuclease with iodoacetic acid leads to the formation of 0.62 residues of pi-carboxymethylhistidine and 0.36 residues of tau-carboxymethylhistidine. The amino acid composition of peptides isolated from diagonal peptide ;maps' and also of peptides isolated after pH1.6 and 2.4 two-dimensional high-voltage electrophoresis of a digest of bison ribonuclease labelled with pyridoxal 5-phosphate indicates that there is complete homology between ox and bison ribonucleases. 4. The Schiff-base attachment site of pyridoxal 5-phosphate was identified as lysine-41 by NaBH(4) reduction followed by peptide isolation.

Citing Articles

Purification and partial characterization of a thiol proteinase from the thermophilic fungus Humicola lanuginosa.

Shenolikar S, Stevenson K Biochem J. 1982; 205(1):147-52.

PMID: 6751320 PMC: 1158457. DOI: 10.1042/bj2050147.

The molecular evolution of pancreatic ribonuclease.

Beintema J, Gaastra W, Lenstra J, Welling G, FITCH W J Mol Evol. 1977; 10(1):49-71.

PMID: 903984 DOI: 10.1007/BF01796134.

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