TY - JOUR
T1 - Testicular proteins associated with the germ cell-marker, TEX101
T2 - Involvement of cellubrevin in TEX101-trafficking to the cell surface during spermatogenesis
AU - Tsukamoto, Hiroki
AU - Yoshitake, Hiroshi
AU - Mori, Miki
AU - Yanagida, Mitsuaki
AU - Takamori, Kenji
AU - Ogawa, Hideoki
AU - Takizawa, Toshihiro
AU - Araki, Yoshihiko
N1 - Funding Information:
The authors are gratefully acknowledged for the staff of the Animal facility, Juntendo University for their technical supports. This work was supported in part by Grants-in-Aid for “High-Tech Research Center” Project for Private Universities: matching fund subsidy and General Scientific Research No. 16591635 and 18591813 from the Ministry of Education, Culture, Sports, Science and Technology, Japan.
PY - 2006/6/23
Y1 - 2006/6/23
N2 - 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.
AB - 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.
KW - Cellubrevin
KW - Germ cell
KW - Glycosylphosphatidylinositol-anchored protein
KW - Mouse
KW - Proteomics
KW - Spermatogenesis
KW - TEX101
KW - Testis
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U2 - 10.1016/j.bbrc.2006.04.070
DO - 10.1016/j.bbrc.2006.04.070
M3 - Article
C2 - 16678124
AN - SCOPUS:33646428686
VL - 345
SP - 229
EP - 238
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -