Flavin-binding sites in Phycomyces

Riboflavin-binding proteins could be the photoreceptors for tropism in the fungus Phycomyces blakesleeanus. Radiolabelled riboflavin bound to both mem brane-associated and cytosolic sites. The membrane sites (approximately 0.2 -0.6 nmol per g fresh material) were highly specific, with decreasing affin ities for riboflavin (K-D approximate to 1 mu M under reducing conditions, K-D approximate to 3 mu M under oxidizing conditions), FMN, roseoflavin, an d FAD. These binding sites, whose properties were similar to those of highe r plants, could be solubilized with mild detergents, and were found in all vegetative parts of the fungus, including the spores. Mutants defective for phototropism did not differ from the wild type in the amount of binding si tes or their affinity. A completely different binding to riboflavin was obs erved in the cytosolic supernatant of the sporangiophores; this activity wa s heat resistant and the binding sites could be partially purified and reco gnized as a polymerization product of gallic acid, Flavins were abundant in the sporangiophores (4.5 nmol per g fresh mass) and the spores (60 nmol pe r g fresh mass), but scarce in washed membranes (0.02-0.11 nmol per g fresh sporangiophore mass). Autogenous fluorescence, whose absorption and emissi on wavelengths fit those of riboflavin, was seen by confocal microscopy, in part as clustered particles, in the actively growing parts of the mycelium , in the cytoplasm of sporangiophores,and in the spores.