An alpha-L-fucosidase potentially involved in fertilization is present on Drosophila spermatozoa surface.
Drosophila is emerging as a model organism to investigate egg fertilization in insects and the possible conservation of molecular mechanisms of gamete interactions demonstrated in higher organisms. This study shows that the spermatozoa of several species of Drosophila belonging to the melanogaster group have a plasma membrane associated alpha-L-fucosidase with features in common with alpha-L-fucosidases from sperm of other animals, including mammals. The enzyme has been purified and completely characterized in D. ananassae, because of its stability in this species. The sperm alpha-L-fucosidase is an integral protein terminally mannosylated, with the catalytic site oriented toward the extracellular space. It has a M(r) of 256 kDa and a multimeric structure made up by subunits of 48 and 55 kDa. Enzyme characterization included kinetic properties, pI, optimal pH, and thermal stability. A soluble form of the enzyme similar to the sperm associated alpha-L-fucosidase is secreted by the seminal vesicles. Synthetic peptides designed from the deduced product of the D. melanogaster gene encoding an alpha-L-fucosidase were used to raise a specific polyclonal antibody. Immunofluorescence labeling of spermatozoa showed that the enzyme is present in the sperm plasma membrane overlying the acrosome and the tail. Lectin cytochemistry analysis of the egg surface indicated that alpha-L-fucose terminal residues are present on the chorion with a strongly polarized localization on the micropyle. The alpha-L-fucosidase of Drosophila sperm plasma membrane appears to be potentially involved in gamete recognition by interacting with its glycoside ligands present on the egg surface at the site of sperm entry.
Intra
Jari
J
Department of Biomolecular Sciences and Biotechnologies, University of Milano, Milano, Italy.
Cenni
Federica
F
Perotti
Maria-Elisa
ME
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology*; D004330:Drosophila / Q000145:classification / Q000166:cytology* / Q000201:enzymology*; D005260:Female; D005306:Fertilization; D006863:Hydrogen-Ion Concentration; D008297:Male; D010063:Ovum / Q000201:enzymology; D011485:Protein Binding; D013094:Spermatozoa / Q000201:enzymology*; D005644:alpha-L-Fucosidase / Q000378:metabolism*
D005644:alpha-L-Fucosidase