Surface of human sperm bears three differently charged CD52 forms, two of which remain stably bound to sperm after capacitation.
gp20 is a sialoglycoprotein of the human sperm surface with a core peptide homologous to the leukocyte antigen CD52, a GPI-anchored glycosylated protein which is described by the monoclonal antibody CAMPATH-1. Comparative analyses, by means of CAMPATH and anti-gp20, indicated that they describe it in morphologically and functionally different ways, suggesting that the respective epitopes are different but also casting doubt on the immunological identity of the antigen. In the present study, we used immunodepletion to demonstrate that CAMPATH and anti-gp20 interact with the same antigen, but that anti-gp20 has a much higher avidity for the antigen than CAMPATH. Anion exchange fractionation analysis of the antigen revealed three differently charged gp20-CD52 forms, the least charged of which, was largely without a GPI-anchor. All three forms were associated with freshly ejaculated sperm, whereas capacitated sperm only contained the two GPI-anchored, more charged forms, which were also the ones found in the prostasome fraction of seminal plasma and in leukocytes. The two charged, GPI-anchored forms were described as homogeneous by anti-gp20, since they ran as a singlet; the third form ran as a doublet. When tested for insertion into Jurkat T cells, the medium charged form inserted the most readily and the less charged one could not be inserted at all.
Della Giovampaola
C
C
Department of Evolutionary Biology, University of Siena, Siena, Italy.
Flori
F
F
Sabatini
L
L
Incerti
L
L
La Sala
G B
GB
Rosati
F
F
Focarelli
R
R
D000918:Antibody Specificity; D015703:Antigens, CD / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; D002457:Cell Extracts; D002462:Cell Membrane / Q000737:chemistry; D002852:Chromatography, Ion Exchange; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D019169:Jurkat Cells; D007962:Leukocytes / Q000737:chemistry / Q000166:cytology; D008297:Male; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D012661:Semen / Q000737:chemistry; D013075:Sperm Capacitation; D013094:Spermatozoa / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D055672:Static Electricity
D015703:Antigens, CD; D000951:Antigens, Neoplasm; D000074301:CD52 Antigen; C000613862:CD52 protein, human; D002457:Cell Extracts; D006023:Glycoproteins; D008562:Membrane Glycoproteins