TY - JOUR
T1 - GalNAcβ1,3-linked paragloboside carries the epitope of a sperm maturation-related glycoprotein that is recognized by the monoclonal antibody MC121
AU - Katagiri, Yohko U.
AU - Sato, Ban
AU - Yamatoya, Kenji
AU - Taki, Takao
AU - Goto-Inoue, Naoko
AU - Setou, Mitsutoshi
AU - Okita, Hajime
AU - Fujimoto, Junichiro
AU - Ito, Chizuru
AU - Toshimori, Kiyotaka
AU - Kiyokawa, Nobutaka
N1 - Funding Information:
We are grateful to Professor Henrik Clausen of University of Copenhagen for providing mAb TH2 and Dr. Kyoko Nakamura for helpful discussion. This work was supported by a grant from the Japan Health Sciences Foundation for Research on Publicly Essential Drugs and Medical Devices ( KHA1002 and KHC1014 ), Health and Labour Sciences Research Grants (the 3rd-term Comprehensive 10-year-strategy for Cancer Control H22-011), and a Grant for Child Health and Development from the Ministry of Health, Labour and Welfare of Japan .
PY - 2011/3/18
Y1 - 2011/3/18
N2 - 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.
AB - 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.
KW - Epididymal maturation
KW - GalNAcβ1-3Gal
KW - MAb MC121
KW - TLC-Blot-MALDI TOF MS
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U2 - 10.1016/j.bbrc.2011.02.019
DO - 10.1016/j.bbrc.2011.02.019
M3 - Article
C2 - 21303663
AN - SCOPUS:79952739335
SN - 0006-291X
VL - 406
SP - 326
EP - 331
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 3
ER -