Expression of senescence-induced protein WS3-10 in vivo and in vitro
Expression of senescence-induced protein WS3-10 in vivo and in vitro
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DOI：
10.1016/0531-5565(95)02015-2
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发表时间：
1996-01-01
影响因子：
3.9
通讯作者：
Goldstein, S
中科院分区：
文献类型：
--
作者：
Grigoriev, VG;Thweatt, R;Goldstein, S
在我们表征细胞衰老的努力中，我们已经表明，与年轻的对应物相比，衰老的人二倍体成纤维细胞（HDF）中编码WS 3 -10蛋白的mRNA过表达，并且年轻HDF中WS 3 -10 cDNA的强制表达导致钙依赖性膜电流的抑制，这可能部分是由于WS 3 -10蛋白内存在钙结合结构域。我们已经在E.大肠杆菌中，并获得亲和纯化的抗体。利用这些抗体的蛋白质印迹分析表明，与年轻HDF相比，衰老HDF中的WS 3 -10蛋白也过表达，与SV 40转化的胎肺HDF相比，正常胎肺HDF中的WS 3 -10蛋白也过表达。HeLa细胞不表达WS 3 -10蛋白，此外，我们在多种大鼠组织中寻找WS 3 -10相关物种。对大鼠组织提取物中的WS 3 -10免疫相关蛋白的分析揭示了两种WS 3 -10同源物，大小为22 kDa和20 kDa。后者可能是由22 kDa多肽的C-末端的蛋白水解去除引起的。这些多肽之间的比例以组织特异性方式变化。在大鼠脾脏和骨骼肌中分别存在大小为39 kDa和91 kDa的与WS 3 -10免疫相关的两种蛋白。
In our efforts to characterize cellular senescence we have shown that the mRNA encoding WS3-10 protein is overexpressed in senescent human diploid fibroblasts (HDF) when compared with their younger counterparts, and that forced expression of the WS3-10 cDNA in young HDF results in suppression of calcium-dependent membrane currents, presumably due in part to the presence of a calcium binding domain within the WS3-10 protein. We have now expressed this protein in E. coli and have obtained affinity purified antibodies. Western blot analysis utilizing these antibodies showed that WS3-10 protein is also overexpressed in senescent HDF when compared to young HDF, and in normal fetal lung HDF when compared to SV40-transformed fetal lung HDF. HeLa cells do not express WS3-10 protein, In addition, we looked for WS3-10-related species in a variety of rat tissues. Analysis of WS3-10 immunologically related proteins in rat tissue extracts revealed two WS3-10 homologs, sized 22 kDa and 20 kDa. The latter presumably result from proteolytic removal of the C-terminal end of the 22 kDa polypeptide. The ratio between these polypeptides varies in a tissue-specific manner. Two proteins immunologically related to WS3-10 with sizes of 39 kDa and 91 kDa were present in rat spleen and skeletal muscle, respectively.