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Immunochemical characterization and localization of nitrate reductase in norflurazon-treated soybean cotyledons
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Immunochemical procedures were used to characterize and localize NADH:nitrate reductase (NR; EC in cotyledons of norflurazon-treated soybeans [Glycine max (L.) Merr. cv. Hill]. Antiserum prepared to NR isolated from Chlorella strongly reacted against NR from norflurazon-treated cotyledons. This serum inhibited the NR activity in crude extracts of norflurazon-treated soybean cotyledons by 98% even at a 1:2000 dilution of crude serum. Pre-immune serum had no effect on the activity. These data indicate that there are similar antigenic determinants at the active site of both Chlorella and norflurazon-treated soybean NR. Whole cotyledons were homogenized in lithium dodecyl sulfate-containing buffer, electrophoretically separated and blotted to nitrocellulose. When the blots were reacted with the anti-NR serum only a single protein (Mr = 98 kdalton) was visualized. Immunofluorescence studies on fixed tissue sections revealed intense fluorescence in the cytoplasm. Weaker reactions were associated with organelles tentatively identified as plastids. Pre-immune serum controls were completely unstained using immunocytochemical procedures. [AS/SS]
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 Record created 1985-06-03, last modified 2019-03-04

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