Hydrophobic properties of phytochrome as probed by 8-anilinonaphthalene-1-sulfonate fluorescence.
8-Anilinonaphthalene-1-sulfonate (ANS) complexes with phytochrome, exhibiting a higher affinity for the Pfr form of phytochrome than for the Pr form. ANS fluorescence is enhanced by the additional binding of ANS to Pfr upon transformation of phytochrome from Pr to Pfr. The specific site of ANS binding appears to be the hydrophobic surface area of the protein, which becomes at least partially exposed in the Pfr form. An exposed, hydrophobic surface area in the Pfr phytochrome has been confirmed by the effects of ANS on the phototransformation of phytochrome. ANS accelerates the Pr leads to Pfr phototransformation, and it inhibits Pfr leads to Pr photoreversion and dark reversion. These effects are interpretable in terms of competitive binding of ANS to the chromophore binding site. Binding of ANS results in a drastic bleaching of the chromophore's absorption bands at 660 and 730 nm, particularly of the latter. This can be attributed to the exposed chromophore, which tends to resume a cyclic conformation with concomitant blue shift and hypochromism of the Qy bands. Sodium dithionite counteracts the inhibitory effects of ANS on the dark reversion of Pfr to Pr, and its effect on the biphasic kinetics of the reversion has been discussed in terms of the Pfr model proposed.
D000817:Anilino Naphthalenesulfonates; D055598:Chemical Phenomena; D002621:Chemistry; D003624:Darkness; D002523:Edible Grain / Q000032:analysis; D007700:Kinetics; D008027:Light; D010834:Phytochrome; D010940:Plant Proteins; D010944:Plants / Q000032:analysis; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D000817:Anilino Naphthalenesulfonates; D010940:Plant Proteins; D010834:Phytochrome