Probing a pheromone binding protein of the silkmoth Antheraea polyphemus by endogenous tryptophan fluorescence.
One subtype of the pheromone binding proteins of the silkmoth Antheraea polyphemus (ApolPBP1) has been analysed exploiting the two endogenous tryptophan residues as fluorescent probe. The intrinsic fluorescence exhibited a rather narrow spectrum with a maximum at 336 nm. Site-directed mutagenesis experiments revealed that one of the tryptophan residues (Trp37) is located in a hydrophobic environment whereas Trp127 is more solvent exposed, as was predicted modeling the ApolPBP1 sequence on the proposed structure of the Bombyx mori pheromone binding protein. Monitoring the interaction of ApolPBP1 as well as its Trp mutants with the three species-specific pheromone compounds by recording the endogenous fluorescence emission revealed profound differences; whereas (E6,Z11)-hexadecadienal induced a dose-dependent quenching of the fluorescence, both (E6,Z11)-hexadecadienyl-1-acetate and (E4,Z9)-tetradecadienyl-1-acetate elicited an augmentation of the endogenous fluorescence. These data indicate that although ApolPBP1 can bind all three pheromones, there are substantial differences concerning their interaction with the protein, which may have important functional implications.
Bette
Stefanie
S
University of Hohenheim, Institute of Physiology, Garbenstrasse 30, 70599, Stuttgart, Germany.
Breer
Heinz
H
Krieger
Jürgen
J
D000085:Acetates / Q000378:metabolism; D000466:Alkadienes / Q000378:metabolism; D000818:Animals; D012831:Bombyx; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D005453:Fluorescence; D019476:Insect Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010675:Pheromones / Q000378:metabolism; D014364:Tryptophan / Q000235:genetics / Q000378:metabolism
C425827:6,11-hexadecadienal; C413925:6,11-hexadecadienyl acetate; D000085:Acetates; D000466:Alkadienes; D002352:Carrier Proteins; D019476:Insect Proteins; D036341:Intercellular Signaling Peptides and Proteins; D010675:Pheromones; C403094:pheromone-binding protein, Bombyx; D014364:Tryptophan