Monoclonal antibodies to a higher-plant nitrate reductase: Differential inhibition of enzyme activities |
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Authors: | B. A. Notton R. J. Fido G. Galfre |
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Affiliation: | (1) Long Ashton Research Station, University of Bristol, Long Ashton, BS18 9AF Bristol;(2) Monoclonal Antibody Centre, Institute of Animal Physiology, Babraham, CB2 4AT Cambridge, UK |
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Abstract: | A set of monoclonal antibodies has been raised against NADH-nitrate reductase (NR; EC 1.6.6.1) from spinach (Spinacea oleracea L.) leaves. Antibodies were screened by enzyme-linked immunosorbent assay and by their ability to inhibit various activities of the enzyme. The six monoclonals selected (AFRC MAC 74 to 79) are all gamma globulins; four (MAC 74 to 77) inhibit all terminal donating activities (NADH-NR; flavin mononucleotide, reduced form (FMNH2)-NR; and methyl viologen, reduced form (MV)-NR) and two (MAC 78 and 79) inhibit the acceptor activities (NADH-NR, and NADH-cytochrome c reductase). MAC 74 to 77 inhibit the NADH-NR activity of crude extracts of a variety of species (mono- and dicotyledoneae) while MAC 78 and 79 are effective against spinach and marrow, but not oil-seed rape, cucumber, oats, wheat and barley.Abbreviations Cyt c Rase cytochrome c reductase - ELISA enzyme-linked immunosorbent assay - FAD(H2) flavin adenine dinucleotide (reduced form) - FMN(H2) flavin mononucleotide (reduced form) - McAb monoclonal antibody - MV methyl viologen reduced form - NR nitrate reductase |
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Keywords: | Immunological discrimination Monoclonal antibody Nitrate reductase Spinacea (nitrate reductase) |
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