Differences between the Surface Properties of the P_R and P_FR Forms of Native Pea Phytochrome, and Their Application to a Simplified Procedure for Puriffication of the Phytochrome

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The ability of phyiochrome from etiolated pea shoots (Pisum sativum L. cv. Alaska) to bind to various chromatographic adsorbents and its mobility during non-denaturing electrophoresis were examined with phytochrome in either the red light-absorbing form (P_R) or the far-red light-absorbing form (P_<FR>). Preferential binding of P_<FR> to modified hydrophilic polyvinyl resins, such as butyl Toyopearl, phenyl Toyopearl, Blue Toyopearl (Cibacron Blue F3G-A conjugated) and Red Toyopearl (Procion Red HE-3B conjugated), was observed. A simplified method for purification of native phyiochrome was developed based on the properties of P_R and P_<FR>. P_<FR> bound preferentially to the hydrophobic adsorbents, to indicate that the surface of P_<FR> is more hydrophobic than that of P_R. A difference in net surface charges between P_R and P_<FR> was detected by an analysis based on the different mobilities of the two forms during non-denaturing polyacrylamide gel electrophoresis in gels prepared with various concentrations of polyacrylamide. The apparent molecular weights of P_R and P_<FR>, estimated from the analysis, were 378 and 419 kilodaltons, respectively. The difference suggests that a significant change in molecular shape occurs during the photoconversion. The differences in surface properties of P_R and P_<FR> are discussed.
日本植物生理学会の論文