Crypticity and functional distribution of the membrane associated alpha-L-fucosidase of human sperm.
Two distinctive isoforms of the enzyme alpha-L-fucosidase are found within human semen in substantial amounts, suggesting specialized functions during reproduction. The membrane-associated isozyme of human sperm cells was previously characterized biochemically, and here we report on its subcellular localization. Intact, detergent permeabilized, capacitated, and acrosome-reacted sperm were investigated using antifucosidase immunofluorescence, binding of the fluorescent fucosylated glycoconjugate RITC-BSA-fucose (RBF), and enzyme activity in the presence and absence of selected inhibitors. Both immunolocalization and RBF binding show that fucosidase is broadly distributed over the membrane systems of human sperm, but is relatively enriched within the equatorial segment. Upon detergent treatment or induction of acrosome reaction (AR), a portion of enzyme activity is recoverable in the supernatant, presumably associated with released remnants of the outer acrosomal membrane. Surprisingly, cell-bound enzyme activity increases sharply following permeabilization of intact sperm, representing cryptic fucosidase that is relatively stable and corresponds with strong fluorescence in the equatorial segment and other sperm membranes. These observations support the notion that the fucosidase has a role in the intimate species signature interactions between sperm and oocyte.
Venditti
Jennifer J
JJ
Department of Biological Sciences, Lehigh University, Bethlehem, Pennsylvania 18015, USA.
Donigan
Katherine A
KA
Bean
Barry S
BS
D000177:Acrosome / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism*; D005643:Fucose / Q000378:metabolism; D006801:Humans; D008297:Male; D011485:Protein Binding; D013075:Sperm Capacitation / Q000502:physiology; D013094:Spermatozoa / Q000201:enzymology / Q000378:metabolism*; D014018:Tissue Distribution; D005644:alpha-L-Fucosidase / Q000378:metabolism*
D005643:Fucose; D005644:alpha-L-Fucosidase