Characterization of fucosyltransferase activity during mouse spermatogenesis: evidence for a cell surface fucosyltransferase.
Fucosyltransferase activity was quantified in mouse germ cells at different stages of spermatogenesis. Specifically, fucosyltransferase activities of pachytene spermatocytes, round spermatids, and cauda epididymal sperm were compared. Fucosyltransferase activity of mixed germ cells displayed an apparent Vmax of 17 pmol (mg of protein)-1 min-1 and an apparent Km of approximately 13 microM for GDP-L-[14C]fucose in the presence of saturating amounts of asialofetuin at 33 degrees C. Under these conditions, cellular fucosyltransferase activity was found to increase during spermatogenesis. In agreement with assays of intact cells, examination of subcellular fractions indicated that a large fraction of fucosyltransferase activity was associated with the cell surface. The fraction of fucosyltransferase activity that was associated with the cell surface progressively increased throughout spermatogenesis and epididymal maturation so that nearly all of the fucosyltransferase in epididymal sperm was on the cell surface. Specifically, by comparison of activities in the presence and absence of the detergent NP-40, the fraction of fucosyltransferase activity that was associated with the cell surface in pachytene spermatocytes, round spermatids, and epididymal sperm was 0.36, 0.5, and 0.85, respectively. These results suggest that a cell surface fucosyltransferase may be important during differentiation of spermatogenic cells in the testis as well as during epididymal maturation and fertilization.
Cardullo
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RA
Department of Anatomy and Cellular Biology, Harvard Medical School, Boston, Massachusetts 02115.
Armant
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DR
Millette
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CF
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D002850:Chromatography, Gel; D005647:Fucosyltransferases / Q000378:metabolism*; D006602:Hexosyltransferases / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D051379:Mice; D008970:Molecular Weight; D013087:Spermatids / Q000201:enzymology; D013090:Spermatocytes / Q000201:enzymology; D013091:Spermatogenesis; D013094:Spermatozoa / Q000201:enzymology* / Q000502:physiology
D005647:Fucosyltransferases; D006602:Hexosyltransferases