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Tritium labelling of ribonuclease by the gas-exposure method, as improved by an electric discharge

✍ Scribed by M. Noyer; A. G. Schnek; J. Léonis; M. Winand

Publisher: John Wiley and Sons
Year: 1976
Tongue: French
Weight: 386 KB
Volume: 12
Category: Article
ISSN: 0022-2135
DOI: 10.1002/jlcr.2580120305

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✦ Synopsis

Abstract

When lyophilized samples of bovine pancreatic ribonuclease are exposed to tritium gas, the exchange process can be markedly improved by applying high frequency electric discharges.

Labelling has been carried out for 5, 15 or 30 minutes periods. Labile radioactivity, was then removed by means of molecular sieving and repeated lyophilizations, the tritiated protein being submitted to further purification by ion‐exchange chromatography.

The specific radioactivity of such purified samples is in the Curie/mmole range, and the biological activity of the enzyme remains well preserved. The distribution of tritium atoms among component aminoacids has been determined, after acid hydrolysis of the labelled protein.

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Optically pure amino-acids were labelled with tritium by the Wilzbach method. The percentage of radioactivity found in the opposite enantiomer was determined by reverse dilution analysis. After tritiation of L-phenylalanine, D-phenylalanine, L-tyrosine, L-proline, D-proline, L-glutamic acid and L-al