Immunological and physical characterization of the products of phytochrome proteolysis

Abstract

The relationship between high molecular weight (large) and low molecular weight (small) forms of phytochrome has been shown earlier to be one of proteolysis. The products of such proteolysis are characterized here by chromatography through Bio-Gel P-200 using specific antiphytochrome sera as an assay system. Degradation of large oat (Avena sativa L. cv. Garry) phytochrome as phytochrome, red-absorbing form, phytochrome, far red-absorbing form, or under cycling conditions in crude preparations containing one or more proteases, always yields one fragment with the immunochemical, electrophoretic, spectroscopic, and size characteristics of small phytochrome. In addition, other fragments are detected which may account, in part, for the different molecular weight estimates reported by others for purified, photoreversible phytochrome. The small phytochrome produced by proteolysis with trypsin of a purified large phytochrome preparation is similar to that produced by the endogenously derived protease(s). A large (estimated molecular weight = 90,000), apparently nonphotoreversible peptide is also identified which is electrophoretically and immunochemically distinct from small phytochrome. Thus, it seems that small phytochrome may not represent more than approximately one-half of the native molecule.