Yoshitake Hiroshi, Tsukamoto Hiroki, Maruyama-Fukushima Mayuko, Takamori Kenji, Ogawa Hideoki, Araki Yoshihiko
Institute for Environmental & Gender-specific Medicine, Juntendo University Graduate School of Medicine, 2-1-1 Tomioka, Urayasu City, Chiba, Japan.
Biochem Biophys Res Commun. 2008 Jul 25;372(2):277-82. doi: 10.1016/j.bbrc.2008.05.088. Epub 2008 May 27.
In adult male mice, the glycosylphosphatidyl inositol-anchored glycoprotein TEX101 is expressed only in germ cells and is thought to be involved in spermatogenesis. However, the details regarding the function of TEX101 remain to be clarified. We previously identified Ly6k as a candidate TEX101-associated protein, but as molecular probes are not currently available to detect Ly6k, we do not have conclusive evidence of the association between TEX101 and Ly6k. In this study, we confirmed the biological interaction between TEX101 and Ly6k using an established anti-mouse Ly6k polyclonal antibody (pAb). A combination of immunoprecipitation, Western blot, and immunohistochemical analyses using the pAb revealed that TEX101 is physically associated with Ly6k within the testis. In addition, these proteins simultaneously co-migrate into the detergent-resistant membrane fractions, suggesting that TEX101 collaborates with Ly6k on the cell membrane and may play a role in spermatogenesis.
在成年雄性小鼠中，糖基磷脂酰肌醇锚定糖蛋白TEX101仅在生殖细胞中表达，被认为参与精子发生。然而，关于TEX101功能的细节仍有待阐明。我们之前将Ly6k鉴定为TEX101相关蛋白的候选者，但由于目前尚无检测Ly6k的分子探针，我们没有TEX101与Ly6k之间关联的确凿证据。在本研究中，我们使用已建立的抗小鼠Ly6k多克隆抗体（pAb）证实了TEX101与Ly6k之间的生物学相互作用。使用该pAb进行免疫沉淀、蛋白质印迹和免疫组织化学分析相结合的结果表明，TEX101在睾丸内与Ly6k存在物理关联。此外，这些蛋白质同时共迁移到抗去污剂膜组分中，表明TEX101与Ly6k在细胞膜上协同作用，可能在精子发生中发挥作用。
