Identification of proteins immunologically related to interferon regulatory factor-1 that bind with interferon regulatory factor element.
Identification of proteins immunologically related to interferon regulatory factor-1 that bind with interferon regulatory factor element.
复制标题
鉴定免疫学上与干扰素调节因子-1 相关且与干扰素调节因子元件结合的蛋白质。
DOI：
10.1086/505358
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发表时间：
2006
期刊：
影响因子：
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通讯作者：
Friedman,RobertM
中科院分区：
文献类型：
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作者：
Contente,Sara;Attard,FrankA;Friedman,RobertM
使用带有 IRF-1 特异性抗血清的蛋白质免疫印迹法，对未转化和癌基因转化的小鼠成纤维细胞中的干扰素调节因子 (IRF)-1 表达进行了调查，以检查细胞转化可能导致的差异。在非转化细胞系和癌基因转化细胞系的提取物中观察到另外十种与 IRF-1 抗体发生反应并与肽抗原进行特异性竞争的蛋白质。在小鼠巨噬细胞提取物中也观察到交叉反应蛋白。使用代表与磁珠连接的 IRF 结合序列的寡核苷酸从成纤维细胞核提取物中捕获蛋白质。使用抗 IRF-1 洗脱捕获的蛋白质并通过免疫印迹进行分析。与 43-kDa IRF-1 一起，7 个位于核的交叉反应蛋白中的 4 个（97、90、66 和 33 kDa）被发现与 IRF 结合元件复合。这些蛋白质具有与 IRF-1 C 末端区域相同的表位和 IRF 元件 DNA 序列结合能力，可能代表 IRF 家族的新成员。
Interferon regulatory factor (IRF)–1 expression was surveyed in nontransformed and oncogene‐transformed mouse fibroblasts, using Western immunoblot with an IRF‐1–specific antiserum, to examine possible differences resulting from cellular transformation. Ten additional proteins that reacted with the IRF‐1 antibody and that underwent specific competition by peptide antigen were observed in extracts of both nontransformed and oncogene‐transformed cell lines. Cross‐reacting proteins were also observed in mouse macrophage extracts. Protein was captured from fibroblast nuclear extracts, using oligonucleotides representing IRF‐binding sequences linked to magnetic beads. Captured proteins were eluted and analyzed by immunoblot with anti–IRF‐1. Along with 43‐kDa IRF‐1, 4 of the 7 nuclearly located cross‐reacting proteins (97, 90, 66, and 33 kDa) were found to complex with the IRF binding element. These proteins, with an epitope in common with the IRF‐1 C‐terminal region and IRF element DNA sequence–binding capability, may represent new members of the IRF family.