Properties of the recombinant TNF-binding proteins from variola, monkeypox, and cowpox viruses are different.
Properties of the recombinant TNF-binding proteins from variola, monkeypox, and cowpox viruses are different.
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DOI：
10.1016/j.bbapap.2006.09.006
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发表时间：
2006-11
期刊：
影响因子：
--
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中科院分区：
文献类型：
--
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肿瘤坏死因子 (TNF) 是一种有效的促炎和抗病毒细胞因子，是痘病毒通过病毒编码的 TNF 结合蛋白家族（CrmB、CrmC、CrmD 和 CrmE）的活性靶向的关键细胞外免疫调节剂。天花病毒 (VARV) 是专门感染人类的天花病原体，其唯一的 TNF 结合蛋白是 CrmB。在这里，我们比对了 10 个天花病毒、14 个牛痘病毒 (CPXV) 和 22 个猴痘病毒 (MPXV) 毒株的 CrmB 蛋白的氨基酸序列。序列分析表明这些蛋白质具有高度同源性。在 CrmB 蛋白序列中发现了与 TNF 受体家族的 cd00185 结构域同源的区域，决定了配体-受体结合的特异性。此外，还对 CrmB 蛋白的 C 端 SECRET 结构域序列进行了比较分析。检测了每个特定正痘病毒物种的这些结构域特征的氨基酸序列的差异。据推测，CrmB 蛋白之间的物种特异性差异可能是这些理化和生物学特性差异的基础。在昆虫细胞的杆状病毒表达系统中合成并分离了各个重组蛋白 VARV-CrmB、MPXV-CrmB 和 CPXV-CrmB。当通过非还原 SDS-PAGE 分级时，纯化的天花病毒-CrmB 可检测为分子量为 90 kDa 的二聚体，而 MPXV-和 CPXV-CrmB 可检测为单体。天花病毒、MPXV 和 CPXV 的 CrmB 蛋白在 L929 小鼠成纤维细胞系中抑制人、小鼠或兔 TNF 细胞毒性作用的效率不同。使用 SPF BALB/c 小鼠在 LPS 诱导休克的实验模型中对 CrmB 进行测试，发现天花病毒 CrmB 具有显着的保护作用。因此，我们的数据证明了 VARV-、MPXV-和 CPXV-CrmB 的抗 TNF 活性的差异，以及 VARV-CrmB 而不是 CPXV- 或 MPXV-CrmB 对 LPS 诱导的小鼠死亡的效率的差异。
Tumor necrosis factor (TNF), a potent proinflammatory and antiviral cytokine, is a critical extracellular immune regulator targeted by poxviruses through the activity of virus-encoded family of TNF-binding proteins (CrmB, CrmC, CrmD, and CrmE). The only TNF-binding protein from variola virus (VARV), the causative agent of smallpox, infecting exclusively humans, is CrmB. Here we have aligned the amino acid sequences of CrmB proteins from 10 VARV, 14 cowpox virus (CPXV), and 22 monkeypox virus (MPXV) strains. Sequence analyses demonstrated a high homology of these proteins. The regions homologous to cd00185 domain of the TNF receptor family, determining the specificity of ligand–receptor binding, were found in the sequences of CrmB proteins. In addition, a comparative analysis of the C-terminal SECRET domain sequences of CrmB proteins was performed. The differences in the amino acid sequences of these domains characteristic of each particular orthopoxvirus species were detected. It was assumed that the species-specific distinctions between the CrmB proteins might underlie the differences in these physicochemical and biological properties. The individual recombinant proteins VARV-CrmB, MPXV-CrmB, and CPXV-CrmB were synthesized in a baculovirus expression system in insect cells and isolated. Purified VARV-CrmB was detectable as a dimer with a molecular weight of 90 kDa, while MPXV- and CPXV-CrmBs, as monomers when fractioned by non-reducing SDS-PAGE. The CrmB proteins of VARV, MPXV, and CPXV differed in the efficiencies of inhibition of the cytotoxic effects of human, mouse, or rabbit TNFs in L929 mouse fibroblast cell line. Testing of CrmBs in the experimental model of LPS-induced shock using SPF BALB/c mice detected a pronounced protective effect of VARV-CrmB. Thus, our data demonstrated the difference in anti-TNF activities of VARV-, MPXV-, and CPXV-CrmBs and efficiency of VARV-CrmB rather than CPXV- or MPXV-CrmBs against LPS-induced mortality in mice.
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DOI：
10.1073/pnas.89.22.10915
发表时间：
1992-11-15
影响因子：
11.1
作者：
HENIKOFF, S;HENIKOFF, JG
通讯作者：
HENIKOFF, JG
影响因子：
12.8
作者：
JACOB, CO
通讯作者：
JACOB, CO
影响因子：
1.2
作者：
Gileva, IP;Ryazankin, IA;Shchelkunov, SN
通讯作者：
Shchelkunov, SN
影响因子：
56.9
作者：
HUANG, S;HENDRIKS, W;AGUET, M
通讯作者：
AGUET, M
影响因子：
4.8
作者：
NAISMITH, JH;DEVINE, TQ;SPRANG, SR
通讯作者：
SPRANG, SR