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Nucleic Acids Research, Vol 26, Issue 18 4306-4307, Copyright © 1998 by Oxford University Press

ARTICLES

A rapid and sensitive method to measure the enzymatic activity of ribosome-inactivating proteins

M Brigotti, L Barbieri, P Valbonesi, F Stirpe, L Montanaro and S Sperti
Dipartimento di Patologia sperimentale dell'Universita degli Studi di Bologna, Via San Giacomo 14, I-40126 Bologna, Italy.

A method is described in which the adenosine- N -glycosidase activity of ribosome-inactivating proteins (RIPs) is measured using as substrate a 2251 bp [3H]DNA obtained by PCR amplification of the 731-2981 region of the pBR322 plasmid. The DNA, labelled in the purine ring of adenine, proved a good substrate for all three RIPs tested (PAP-S, ricin and shiga-like toxin I). The method, which measures directly the [3H]adenine released, is highly specific, extremely rapid and quantitative in a wide range of RIP concentrations.
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