Neu-Yilik G, Zorbas H, Gloe T R, Raabe H M, Hopp-Christensen T A, Müller P K
Medizinische Universität zu Lübeck, Institut für Med. Molekularbiologie, Germany.
Eur J Biochem. 1993 Apr 15;213(2):727-36. doi: 10.1111/j.1432-1033.1993.tb17813.x.
A fusion protein composed of about two vigilin domains and beta-galactosidase was used to raise polyclonal antibodies which were affinity-purified and employed for immunoblotting and immunohistochemistry. A protein of an apparent molecular mass of 155 kDa could be stained in extracts of a variety of cells from different species and organs. Immunohistological studies on single cells showed that vigilin is accumulated in the cytoplasm. During in vitro maintenance of primary cell cultures, as well as of a growth-factor-dependent cell line, vigilin expression decreases and ceases in senescent cells. In contrast, vigilin is constitutively expressed in all other transformed cell lines of various origin studies so far. Vigilin expression can be induced in peripheral blood lymphocytes by mitogen stimulation. These observations suggest an involvement of vigilin in processes of cell activation. Immunoblot experiments demonstrating the presence of vigilin in a broad range of eukaryotes, indicate a high degree of evolutionary conservation.
一种由大约两个维吉琳结构域和β-半乳糖苷酶组成的融合蛋白被用于制备多克隆抗体，这些抗体经过亲和纯化后用于免疫印迹和免疫组织化学。在来自不同物种和器官的多种细胞提取物中，可以检测到一种表观分子量为155 kDa的蛋白质。单细胞免疫组织学研究表明，维吉琳在细胞质中积累。在原代细胞培养以及生长因子依赖性细胞系的体外维持过程中，维吉琳的表达在衰老细胞中会降低并停止。相比之下，在迄今为止研究的各种来源的所有其他转化细胞系中，维吉琳都是组成性表达的。有丝分裂原刺激可诱导外周血淋巴细胞中维吉琳的表达。这些观察结果表明维吉琳参与细胞激活过程。免疫印迹实验证明维吉琳存在于广泛的真核生物中，这表明其具有高度的进化保守性。
