Abstract
Riboflavin-binding proteins could be the photoreceptors for tropism in the fungus
Phycomyces blakesleeanus. Radiolabelled riboflavin bound to both membrane-associated and cytosolic sites.
The membrane sites (approximately 0.2-0.6 nmol per g fresh material) were highly specific,
with decreasing affinities for riboflavin (KD ≈ 1 µM under reducing conditions, KD ≈ 3 µM under oxidizing conditions), FMN, roseoflavin, and FAD. These binding sites,
whose properties were similar to those of higher 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 sites or their affinity. A completely different binding to riboflavin
was observed in the cytosolic supernatant of the sporangiophores; this activity was
heat resistant and the binding sites could be partially purified and recognized 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 per g fresh mass), but scarce
in washed membranes (0.02-0.11 nmol per g fresh sporangiophore mass). Autogenous fluorescence,
whose absorption and emission 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.
Key words
Blue light receptors - gallic acid - membranes -
Phycomyces
- riboflavin
