title: GalNAcβ1,3-linked paragloboside carries the epitope of a sperm maturation-related glycoprotein that is recognized by the monoclonal antibody MC121.
abstract: The functional maturation of spermatozoa during epididymal transit in mammals accompanies the changes in their plasma membrane due to the binding or removal of proteins or interactions with the proteases, glycosidases and glycosyltransferases present in the epididymis. In order to study the surface changes in spermatozoa during their maturation in the epididymis, we previously established several monoclonal antibodies against the 54kDa sialoglycoprotein of mouse cauda epididymal spermatozoa, which gradually increased the expression of antigenic determinants during epididymal transit. One of these monoclonal antibodies, MC121, reacted with mouse sperm glycoproteins on a polyvinylidene fluoride membrane after desialylation of the glycoproteins, and the treatment of the desialylated sperm glycoproteins with β-N-acetylhexosaminidase greatly decreased the expression of the antigenic determinants. In addition to reacting with mouse cauda epididymal spermatozoa, MC121 reacted with human red blood cells (hRBCs). MC121 induced agglutination of sialidase-treated hRBCs and stained hRBCs fixed with formalin vapor much more heavily than it stained hRBCs fixed with methanol. The thin layer chromatography (TLC) immunostaining of the sialidase-treated lipids of hRBCs with MC121 suggested that the epitope-bearing molecule is a glycosphingolipids (GSL), and that MC121 reacts with a pentaose-GSL. Analysis of sialidase-treated GSLs by TLC-Blot-Matrix Assisted Laser Desorption Ionization Time-of-Flight mass spectrometry (MALDI TOF MS) revealed that the GSL bound by MC121 was [HexNAc][HexNAc+Hex][Hex][Hex]-Cer. The lipid band stained with mAb TH2, which is specific for a GSL, GalNAcβ1-3Galβ1-4GlcNAcβ1-3Galβ1-4Glcβ1-ceramide. These results indicated that the epitope to which MC121 binds is present in a neolacto-series GSL, IV³GalNAcβ-nLc₄Cer² sequence.
authors.affiliation: Dept. of Pediatric Hematology and Oncology, Natl. Res. Inst. for Child Health & Development, Tokyo 157-8535, Japan. kata@nch.go.jp
authors.forename: Yohko U
authors.initials: YU
authors.lastname: Katagiri
authors.forename: Ban
authors.initials: B
authors.lastname: Sato
authors.forename: Kenji
authors.initials: K
authors.lastname: Yamatoya
authors.forename: Takao
authors.initials: T
authors.lastname: Taki
authors.forename: Naoko
authors.initials: N
authors.lastname: Goto-Inoue
authors.forename: Mitsutoshi
authors.initials: M
authors.lastname: Setou
authors.forename: Hajime
authors.initials: H
authors.lastname: Okita
authors.forename: Junichiro
authors.initials: J
authors.lastname: Fujimoto
authors.forename: Chizuru
authors.initials: C
authors.lastname: Ito
authors.forename: Kiyotaka
authors.initials: K
authors.lastname: Toshimori
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Kiyokawa
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000937:Antigen-Antibody Reactions; D002240:Carbohydrate Sequence; D004822:Epididymis / Q000276:immunology*; D005915:Globosides / Q000737:chemistry / Q000276:immunology*; D016056:Immunodominant Epitopes / Q000276:immunology*; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D012795:Sialoglycoproteins / Q000737:chemistry / Q000276:immunology*; D013079:Sperm Maturation / Q000276:immunology*; D013082:Sperm Tail / Q000276:immunology*; D054821:beta-Hexosaminidase beta Chain / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D005915:Globosides; D016056:Immunodominant Epitopes; D012795:Sialoglycoproteins; C036363:paragloboside; D054821:beta-Hexosaminidase beta Chain