Plant Physiol. Drug Metab Dispos
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Plant Physiology 79:62-64 (1985)
© 1985 American Society of Plant Biologists

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Articles

Determination of Diamine Oxidase in Lentil Seedlings by Enzymic Activity and Immunoreactivity

Rodolfo Federico, Riccardo Angelini, Alberinda Cesta and Carlo Pini

Dipartimento di Biologia Vegetale, Università "La Sapienza," P. le Aldo Moro 5, 00185 Rome, Italy, Istituto Superiore di Sanità, Vl. Regina Elena 299, 00185 Rome, Italy

A competitive radioimmunoassay for the quantitation of diamine oxidase (EC 1.4.3.6) from Lens culinaris is reported. Specific antibodies raised in rabbits immunized with a homogeneous preparation of the enzyme were incubated with purified 125I-enzyme and with either unlabeled diamine oxidase or plant material. Antigen-antibody complexes were isolated from the mixture by incubation with Staphylococcus protein A. The sensitivity of the test was about 5 nanograms in terms of enzyme protein. This assay was applied to the determination of the enzyme in extracts from lentil shoots grown either in the dark or in the light. Diamine oxidase activity and enzyme protein (as determined by radioimmunoassay) were measured during 7 days after germination. Both enzymic activity and enzyme protein declined slowly in the dark and rapidly in the light. These results indicate that fluctuation of the enzymic activity in this organ, both in the light and in the dark, are mediated via changes in the amount of the enzyme protein and not via the action of an inhibitor.




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Copyright © 1985 by the American Society of Plant Biologists