Hostname: page-component-cd9895bd7-gvvz8 Total loading time: 0 Render date: 2024-12-27T07:58:11.918Z Has data issue: false hasContentIssue false
  • English
  • Français

Molecular characterization of a germ-cell-specific antigen, TEX101, from mouse testis

Published online by Cambridge University Press:  01 August 2006

Hong Jin ,
Hiroshi Yoshitake ,
Hiroki Tsukamoto ,
Mai Takahashi ,
Miki Mori ,
Toshihiro Takizawa ,
Kenji Takamori ,
Hideoki Ogawa ,
Katsuyuki Kinoshita  and
Yoshihiko Araki
Hong Jin
Affiliation:
Institute for Environmental & Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba 279-0021, Japan. Department of Obstetrics & Gynecology, Juntendo University Faculty of Medicine, Tokyo 113-8595, Japan.
Hiroshi Yoshitake
Affiliation:
Institute for Environmental & Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba 279-0021, Japan.
Hiroki Tsukamoto
Affiliation:
Institute for Environmental & Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba 279-0021, Japan.
Mai Takahashi
Affiliation:
Institute for Environmental & Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba 279-0021, Japan.
Miki Mori
Affiliation:
Department of Molecular Anatomy, Nippon Medical School, Tokyo 113-8602, Japan.
Toshihiro Takizawa
Affiliation:
Department of Molecular Anatomy, Nippon Medical School, Tokyo 113-8602, Japan.
Kenji Takamori
Affiliation:
Institute for Environmental & Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba 279-0021, Japan.
Hideoki Ogawa
Affiliation:
Institute for Environmental & Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba 279-0021, Japan.
Katsuyuki Kinoshita
Affiliation:
Department of Obstetrics & Gynecology, Juntendo University Faculty of Medicine, Tokyo 113-8595, Japan.
Yoshihiko Araki*
Affiliation:
Institute for Environmental & Gender-Specific Medicine, Juntendo University Graduate School of Medicine, Chiba 279-0021, Japan.
*
All correspondence to: Yoshihiko Araki, MD, DMedSci, Institute for Environmental and Gender-Specific Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Tomioka, Urayasu-City, Chiba 279-0021, Japan. Fax: +81 47 3533178. e-mail: yaraki@med.juntendo.ac.jp
Get access Rights & Permissions [Opens in a new window]

Summary

TEX101, a glycoprotein we recently identified, is primarily characterized as a unique germ-cell-specific marker protein that shows sexually dimorphic expression during mouse gonad development. Based on data obtained from molecular biological as well as immuno-morphological studies, we believe this molecule may play a role in the process underlying germ cell formation. However, many points remain unclear as the molecular characteristics and its physiological functions are far from being completely understood. To clarify the molecular basis of TEX101, we herein report a further biochemical characterization of the molecule using testicular Triton X-100 extracts from mice. Deglycosylation studies using endoglycohydrolases that delete N-linked oligosaccharides (OS) from the molecule show that TEX101 is highly (approximately 47%) N-glycosylated. All potential N-glycosylation sites within TEX101 are glycosylated and most of these sites are occupied by endoglycosidase F2-sensitive biantennary complex type OS units. In addition, an extremely low population among TEX101 possesses only endoglycosidase H-sensitive hybrid type OS units. In studies using phosphatidylinositol-specific phospholipase C against native testicular cells or TEX101 transfectant, the enzyme treatment caused major reduction of the TEX101 expression on the cell, suggesting that TEX101, at least in part, is expressed as a glycosylphosphatidylinositol-anchored protein. Taken together, these findings will help elucidate the molecular nature of TEX101, a marker molecule that appeared on germ cells during gametogenesis.

Keywords

GPI-anchored proteinGlycoproteinSpermatogenesisTestisTEX101
Type
Research Article
Information
Zygote , Volume 14 , Issue 3 , August 2006 , pp. 201 - 208
Copyright
Copyright © Cambridge University Press 2006

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Anzai-Takeda, Y., Takeda, T., Sendo, F. & Araki, Y. (2005). Inhibition of cell spreading and stress fiber formation in CHO cells transfected with cDNA of a glycosylphosphatidylinositol-anchored glycoprotein, GPI-80. Immunobiology 210, 1–10.CrossRefGoogle Scholar
Araki, Y., Orgebin-Crist, M.-C. & Tulsiani, D.R.P. (1992 a). Qualitative characterization of oligosaccharide chains present on the rat zona pellucida glycoconjugates. Biol. Reprod. 46, 912–19.CrossRefGoogle ScholarPubMed
Araki, Y., Vierula, M.E., Rankin, T.L., Tulsiani, D.R.P. & Orgebin-Crist, M.-C. (1992 b). Isolation and characterization of a 25-kilodalton protein from mouse testis: sequence homology with a phospholipid-binding protein. Biol. Reprod. 47, 832–43.CrossRefGoogle ScholarPubMed
Barsumian, E.L., Isersky, C., Petrino, M.G. & Siraganian, R.P. (1981). IgE-induced histamine release from rat basophilic leukemia cell lines: isolation of releasing and nonreleasing clones. Eur. J. Immunol. 11, 317–23.CrossRefGoogle ScholarPubMed
Graham, F.L., Smiley, J., Russell, W.C. & Nairn, R. (1977). Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J. Gen. Virol. 36, 59–74.CrossRefGoogle ScholarPubMed
Halova, I., Draberova, L. & Draber, P. (2002). A novel lipid raft-associated glycoprotein, TEC-21, activates rat basophilic leukemia cells independently of the type 1 Fcϵ receptor. Int. Immunol. 14, 213–23.CrossRefGoogle Scholar
Kinoshita, T., Ohishi, K. & Takeda, J. (1997). GPI-anchor synthesis in mammalian cells: genes, their products, and a deficiency. J. Biochem. 122, 251–7.CrossRefGoogle ScholarPubMed
Kurita, A., Takizawa, T., Takayama, T., Totsukawa, K., Matsubara, S., Shibahara, H., Orgebin-Crist, M.-C., Sendo, F., Shinkai, Y. & Araki, Y. (2001). Identification, cloning, and initial characterization of a novel mouse testicular germ cell-specific antigen. Biol. Reprod. 64, 935–45.CrossRefGoogle ScholarPubMed
Laemmli, U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–5.CrossRefGoogle ScholarPubMed
Ohtake, K., Takei, H., Watanabe, T., Sato, Y., Yamashita, T., Sudo, K., Kuroki, M., Chihara, J. & Sendo, F. (1997). A monoclonal antibody modulates neutrophil adherence while enhancing cell motility. Microbiol. Immunol. 41, 67–72.CrossRefGoogle ScholarPubMed
Plummer, T.H. Jr & Tarentino, A.L. (1991). Purification of the oligosaccharide-cleaving enzymes of Flavobacterium meningosepticum. Glycobiology 1, 257–63.CrossRefGoogle ScholarPubMed
Sayama, K., Shiraishi, S., Shirakata, Y., Kobayashi, Y. & Miki, Y. (1991). Characterization of decay-accelerating factor (DAF). in human skin. J. Invest. Dermatol. 96, 61–4.CrossRefGoogle ScholarPubMed
Sleight, S.B., Miranda, P.V., Plaskett, N.W., Maier, B., Lysiak, J., Scrable, H., Herr, J.C. & Visconti, P.E. (2005). Isolation and proteomic analysis of mouse sperm detergent-resistant membrane fractions: evidence for dissociation of lipid rafts during capacitation. Biol. Reprod. 73, 721–9.CrossRefGoogle ScholarPubMed
Suzuki, K., Watanabe, T., Sakurai, S., Ohtake, K., Kinoshita, T., Araki, A., Fujita, T., Takei, H., Takeda, Y., Sato, Y., Yamashita, T., Araki, Y. & Sendo, F. (1999). A novel glycosylphosphatidyl inositol-anchored protein on human leukocytes: a possible role for regulation of neutrophil adherence and migration. J. Immunol. 162, 4277–84.CrossRefGoogle ScholarPubMed
Takayama, T., Mishima, T., Mori, M., Jin, H., Tsukamoto, H., Takahashi, K., Kinoshita, K., Suzuki, M., Sato, I., Matsubara, S., Araki, Y. & Takizawa, T. (2005 a). Sexually dimorphic expression of the novel germ cell antigen TEX101 during mouse gonad development. Biol. Reprod. 72, 1315–23.CrossRefGoogle ScholarPubMed
Takayama, T., Mishima, T., Mori, M., Ishikawa, T., Takizawa, T., Goto, T., Suzuki, M., Araki, Y., Matsubara, S. & Takizawa, T. (2005 b). TEX101 is shed from the surface of sperm located in the caput epididymidis of the mouse. Zygote 13, 325–333.CrossRefGoogle ScholarPubMed
Tarentino, A.L. & Maley, F. (1974). Purification and properties of an end-β-N-acetylglucosaminidase from Streptomyces griseus. J. Biol. Chem. 249, 811–17.CrossRefGoogle Scholar
Tarentino, A.L., Gomez, C.M. & Plummer, T.H. Jr (1985). Deglycosylation of asparagine-linked glycans by peptide: N-glycosidase F. Biochemistry 24, 4665–71.CrossRefGoogle ScholarPubMed
Trimble, R.B. & Tarentino, A.L. (1991). Identification of distinct endoglycosidase (endo) activities in Flavobacterium meningosepticum: endo F1, endo F2, and endo F3. Endo F1 and endo H hydrolyze only high mannose and hybrid glycans. J. Biol. Chem. 266, 1646–51.CrossRefGoogle Scholar
Tsukamoto, H., Takizawa, T., Takamori, K., Ogawa, H. & Araki, Y. (2006). Genomic organization and structure of the 5′-flanking region of the TEX101 gene: alternative promo-ter usage and splicing generate transcript variants with distinct 5′-untranslated region. Mol. Reprod. Dev. (in press).Google Scholar
Udenfriend, S. & Kodukula, K. (1995). How glycosylphosphatidylinositol-anchored membrane proteins are made. Annu. Rev. Biochem. 64, 563–91.CrossRefGoogle Scholar
Yeh, E.T.H., Kamitani, T. & Chang, H.M. (1994). Biosynthesis and processing of the glycosylphosphatidylinositol anchor in mammalian cells. In Seminar in Immunology, vol. 6 (ed. Ashwell, J.D.), pp. 73–80. London: Academic Press.Google Scholar
Yu, Y., Araki, Y. & Sendo, F. (2000). Tyrosine phosphorylation of a 34-kDa protein induced by crosslinking of a novel glycosylphosphatidylinositol-anchored glycoprotein (GPI-80). on human neutrophils that may regulate their adherence and migration. IUBMB Life 49, 43–7.Google ScholarPubMed