GalNAcβ1,3-linked paragloboside carries the epitope of a sperm maturation-related glycoprotein that is recognized by the monoclonal antibody MC121

Yohko U. Katagiri, Ban Sato, Kenji Yamatoya, Takao Taki, Naoko Goto-Inoue, Mitsutoshi Setou, Hajime Okita, Junichiro Fujimoto, Chizuru Ito, Kiyotaka Toshimori, Nobutaka Kiyokawa

Research output: Contribution to journalArticle

7 Citations (Scopus)

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-nLc4Cer2The nomenclature for glycosphingolipids follows the recommendations [26] of the IUPAC-IUB, and the ganglioside nomenclature of Svennerholm [27] was used.2 sequence.

Original languageEnglish
Pages (from-to)326-331
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume406
Issue number3
DOIs
Publication statusPublished - 2011 Mar 18
Externally publishedYes

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Sperm Maturation
Glycosphingolipids
Epitopes
Glycoproteins
Monoclonal Antibodies
Spermatozoa
Blood
Erythrocytes
Neuraminidase
Thin layer chromatography
Epididymis
Terminology
Thin Layer Chromatography
Bearings (structural)
Asialoglycoproteins
Cells
Sialoglycoproteins
Lipids
Glycosyltransferases
Mammals

Keywords

  • Epididymal maturation
  • GalNAcβ1-3Gal
  • MAb MC121
  • TLC-Blot-MALDI TOF MS

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

GalNAcβ1,3-linked paragloboside carries the epitope of a sperm maturation-related glycoprotein that is recognized by the monoclonal antibody MC121. / Katagiri, Yohko U.; Sato, Ban; Yamatoya, Kenji; Taki, Takao; Goto-Inoue, Naoko; Setou, Mitsutoshi; Okita, Hajime; Fujimoto, Junichiro; Ito, Chizuru; Toshimori, Kiyotaka; Kiyokawa, Nobutaka.

In: Biochemical and Biophysical Research Communications, Vol. 406, No. 3, 18.03.2011, p. 326-331.

Research output: Contribution to journalArticle

Katagiri, Yohko U. ; Sato, Ban ; Yamatoya, Kenji ; Taki, Takao ; Goto-Inoue, Naoko ; Setou, Mitsutoshi ; Okita, Hajime ; Fujimoto, Junichiro ; Ito, Chizuru ; Toshimori, Kiyotaka ; Kiyokawa, Nobutaka. / GalNAcβ1,3-linked paragloboside carries the epitope of a sperm maturation-related glycoprotein that is recognized by the monoclonal antibody MC121. In: Biochemical and Biophysical Research Communications. 2011 ; Vol. 406, No. 3. pp. 326-331.
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AU - Taki, Takao

AU - Goto-Inoue, Naoko

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AU - Okita, Hajime

AU - Fujimoto, Junichiro

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