Testicular proteins associated with the germ cell-marker, TEX101: Involvement of cellubrevin in TEX101-trafficking to the cell surface during spermatogenesis

Hiroki Tsukamoto, Hiroshi Yoshitake, Miki Mori, Mitsuaki Yanagida, Kenji Takamori, Hideoki Ogawa, Toshihiro Takizawa, Yoshihiko Araki

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Recently, we identified a cell-surface marker protein, TEX101, that is unique to male and female germ cells. On/off switching of TEX101 expression in germ cells is closely linked to the kinetics of gametogenesis. In the present study, we isolated testicular proteins by immunoprecipitation with anti-TEX101 antibody and identified the proteins using liquid chromatography/tandem mass spectrometry. Of three proteins identified (annexin 2, ly6k, and cellubrevin), a biochemical association between TEX101 and cellubrevin was confirmed by immunoprecipitation-Western blotting experiments. Immunohistochemistry using a cellubrevin-specific antibody indicated that the molecule is abundant on spermatocytes and early-stage spermatids, whereas negligible amounts are found in Sertoli cells, spermatogonia, spermatozoa, and late-stage spermatids. Most of the intracellular cellubrevin appeared to be juxtaposed with intracellular TEX101, and membrane-associated cellubrevin was docked near TEX101-positive plasma membranes on the cytoplasmic side. This close association was never observed on the outer surface of the plasma membrane. From these results we concluded that cellubrevin-dependent membrane trafficking is involved in TEX101-transport to the surface of male germ cells.

Original languageEnglish
Pages (from-to)229-238
Number of pages10
JournalBiochemical and biophysical research communications
Volume345
Issue number1
DOIs
Publication statusPublished - 2006 Jun 23

Keywords

  • Cellubrevin
  • Germ cell
  • Glycosylphosphatidylinositol-anchored protein
  • Mouse
  • Proteomics
  • Spermatogenesis
  • TEX101
  • Testis

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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