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Separation of Particulate Phytochrome and Plasma Membranes of Maize Coleoptiles by Density Gradient Centrifugation
Authors:YU, R.   CARTER, J.   OSAWA, T.
Abstract:Phytochrome, cross linked in situ to its receptor by glutaraldehydefixation, and radioactively-labelled membrane material, obtainedby lactoperoxidase-catalysed 125I-treatment of maize coleoptilescould be separated from each other according to sedimentationvelocity and by sucrose density gradient centrifugation. Bindingof iodine to membrane material in maize coleoptiles increasedseveral-fold on the addition of reaction-specific enzymes, i.e.lactoperoxidase and glucose oxidase. Mitochondria were considerednot labelled because mito-chondrial purification reduced 125Iincorporation. Membrane material containing incorporated iodineappeared quite heterogenous and sedimented to an equilibriumdensity position close to, but slightly lighter than that ofthe mitochondria; participate phytochrome banded at a heavierposition. Results obtained therefore suggest that the receptorfor phytochrome may not be on the plasma membrane as envisagedin recent hypotheses.
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