HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 68/3/709    most recent biolreprod.102.004465v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Khunsook, S. Articles by Alhadeff, J. A. Search for Related Content PubMed PubMed Citation Articles by Khunsook, S. Articles by Alhadeff, J. A. Agricola Articles by Khunsook, S. Articles by Alhadeff, J. A.

Purification and Characterization of Plasma Membrane-Associated Human Sperm -L-Fucosidase¹

^a Department of Biological Sciences ^b Division of Biochemical Sciences, Department of Chemistry, Lehigh University, Bethlehem, Pennsylvania 18015

Detergent and salt extraction studies, as well as cytochemical localization with fluorescein isothiocyanate-bovine serum albumin-L-fucose, have provided further evidence for the plasma membrane association of a novel human sperm, -L-fucosidase. This -L-fucosidase has been solubilized and purified 8600-fold to high specific activity (35 000 U/mg protein) by affinity chromatography on agarose-C₂₄-fucosylamine. To our knowledge, this is the first report concerning the purification and characterization of a mammalian plasma membrane-associated -L-fucosidase. Both SDS-PAGE and Western blot analysis indicated the -L-fucosidase is highly purified and contains a single subunit with a molecular mass of 51 kDa. N-glycanase studies indicated the subunit contains N-glycans, and lectin blot analysis detected the presence of mannose, but no terminal galactose or sialic acid residues. Isoelectric focusing indicated the presence of two major -L-fucosidase isoforms (pIs 6.5 and 6.7) and a possible minor isoform (pI 6.3). Treatment of -L-fucosidase with neuraminidase did not change its isoform profile, providing further evidence for the enzyme's lack of sialic acid residues. Kinetic analysis with 4-methylumbelliferyl -L-fucopyranoside indicated that sperm -L-fucosidase has a pH optimum near 7, an apparent K_m of 0.08 mM, and a V_max of 6.8 µmol/min/mg protein. The unusual properties of human sperm -L-fucosidase argue in support of a potentially important, but presently unknown, role for this enzyme in human reproduction.

¹ Supported in part by a grant for Undergraduate Education in Biological Sciences from the Howard Hughes Medical Institute. S.K. was supported as a scholar of the Thai Government Ministry of University Affairs.

² Correspondence: Barry Bean, Lehigh University, 111 Research Drive, Bethlehem, PA 18015. FAX: 610 758 4004; e-mail: bb00{at}lehigh.edu

³ Current address: Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen, Thailand 40002

This article has been cited by other articles:

				P. C. N. Chiu, M.-K. Chung, R. Koistinen, H. Koistinen, M. Seppala, P.-C. Ho, E. H. Y. Ng, K.-F. Lee, and W. S. B. Yeung Glycodelin-A interacts with fucosyltransferase on human sperm plasma membrane to inhibit spermatozoa-zona pellucida binding J. Cell Sci., January 1, 2007; 120(1): 33 - 44. [Abstract] [Full Text] [PDF]

				F. Cattaneo, M. E. Pasini, J. Intra, M. Matsumoto, F. Briani, M. Hoshi, and M. E. Perotti Identification and expression analysis of Drosophila melanogaster genes encoding {beta}-hexosaminidases of the sperm plasma membrane Glycobiology, September 1, 2006; 16(9): 786 - 800. [Abstract] [Full Text] [PDF]

				P.C.N. Chiu, H.Y. Tsang, R. Koistinen, H. Koistinen, M. Seppala, K.F. Lee, and W.S.B. Yeung The Contribution of D-Mannose, L-Fucose, N-Acetylglucosamine, and Selectin Residues on the Binding of Glycodelin Isoforms to Human Spermatozoa Biol Reprod, June 1, 2004; 70(6): 1710 - 1719. [Abstract] [Full Text] [PDF]