Sorcin (V19), a soluble acidic calcium-binding protein overproduced in multidrug-resistant cells. Identification of the protein by anti-sorcin antibody.
Sorcin (V19), a soluble acidic calcium-binding protein overproduced in multidrug-resistant cells. Identification of the protein by anti-sorcin antibody.
复制标题
DOI：
10.1016/0006-2952(87)90606-x
复制
发表时间：
1987-07
影响因子：
5.8
通讯作者：
M. B. Meyers;K. Schneider;B. Spengler;T. D. Chang;J. L. Biedler
中科院分区：
文献类型：
--
作者：
M. B. Meyers;K. Schneider;B. Spengler;T. D. Chang;J. L. Biedler
Sorcin（可溶性耐药相关钙结合蛋白）是一种酸性 (pI = 5.7) 蛋白 (Mr∼ 20 kDa)，以前称为 V19，最初是在选择对长春新碱具有高水平耐药性的细胞中发现的。二维凝胶电泳和/或蛋白质印迹技术现在显示，在选择的抗放线菌素 D (QUA/ADj)、秋水仙碱 (CHRC5) 和阿霉素 (BE(2)-C/ADR) 抗性的细胞中，sorcin 过量产生。并非所有选择对这些药物具有抗性的细胞系都会过量产生 sorcin；例如独立选择的 QUA 耐放线菌亚系 QUA/ADsx 的细胞不含有增加量的 sorcin。通过制备型凝胶电泳从 QUA/ADj 细胞中纯化 Sorcin，并用于在鸡中产生特异性抗血清。通过蛋白质印迹分析，显示抗血清可识别 QUA/ADj 以及长春新碱抗性小鼠和中国仓鼠肺、秋水仙碱抗性中国仓鼠卵巢和阿霉素抗性人神经母细胞瘤系中的 sorcin。在对照药物敏感细胞中可检测到该蛋白的低水平表达。与 45Ca2+ 的直接结合测定表明，sorcin 是一种钙结合蛋白。 QUA/ADj 细胞含有数量增加的双微小染色体 (DM)，这是基因扩增的细胞遗传学指标。正如在另外两个多重耐药亚系中所发现的，QUA/ADj 细胞中 sorcin 过量产生可能是 sorcin 编码基因扩增的结果。该蛋白在不同物种的多重耐药细胞中的过量产生表明 sorcin 在耐药表型的表达中发挥作用。
Sorcin (soluble resistance-related calcium-binding protein), an acidic (pI = 5.7) protein (Mr∼ 20 kDa) previously designated V19, was originally identified in cells selected for high levels of resistance to vincristine. Two-dimensional gel electrophoresis and/or Western blot techniques now show sorcin to be overproduced in cells selected for resistance to actinomycin D (QUA/ADj), colchicine (CHRC5), and adriamycin (BE(2)-C/ADR). Not all cell lines selected for resistance to these drugs overproduced sorcin; e.g. cells of an independently selected actinomycind-resistant subline of QUA, QUA/ADsx, did not contain increased amounts of sorcin. Sorcin was purified by preparative gel electrophoresis from QUA/ADj cells and used to generate specific antiserum in chickens. By Western blot analyses the antiserum was shown to recognize sorcin in QUA/ADj and in vincristine-resistant mouse and Chinese hamster lung, colchicine-resistant Chinese hamster ovary, and adriamycin-resistant human neuroblastoma lines. Low level expression of the protein was detectable in control, drug-sensitive cells. Direct binding assays with45Ca2+showed that sorcin was a calcium-binding protein. QUA/ADj cells contained increased numbers of double minute chromosomes (DMs), cytogenetic indicators of gene amplification. As found for two other multidrug-resistant sublines, sorcin overproduction in QUA/ ADj cells may be the result of amplification of the sorcin-encoding gene. The overproduction of this protein in multidrug-resistant cells of various species implies that sorcin plays a role in expression of the resistant phenotype.