Heat shock protein 70 in lung and kidney of specific-pathogen-free chickens is a receptor-associated protein that interacts with the binding domain of the spike protein of infectious bronchitis virus.
Heat shock protein 70 in lung and kidney of specific-pathogen-free chickens is a receptor-associated protein that interacts with the binding domain of the spike protein of infectious bronchitis virus.
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无特定病原体鸡肺和肾中的热休克蛋白70是一种受体相关蛋白，与传染性支气管炎病毒刺突蛋白的结合域相互作用
DOI：
10.1007/s00705-017-3280-x
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发表时间：
2017-06
影响因子：
2.7
通讯作者：
Wang H
中科院分区：
文献类型：
--
作者：
Zhang Z;Yang X;Xu P;Wu X;Zhou L;Wang H
鸡传染性支气管炎病毒（IBV）是冠状病毒科（Coronaviridae）的成员。在IBV原型毒株M41的S1蛋白中已经鉴定出介导病毒与其受体附着的结合结构域。首先，我们表达了M41和SCZJ 3的S1 N蛋白（残基19-270），并比较了重组S1 N-M41和S1 N-SCZJ 3与宿主组织的结合能力。蛋白质组织化学显示S1 N-M41和S1 N-SCZJ 3都可以与肺和肾脏结合，并且重组S1 N-SCZJ 3在前胃中显示出独特的染色模式。重组S1 N-SCZJ 3然后被用于纯化肺、肾和腺胃中的结合相关蛋白。使用亲和层析测定，从总组织蛋白中获得约60 kDa和70 kDa的两个共同条带。这些蛋白条带通过液相色谱-串联质谱（LC-MS/MS）鉴定为蛋白质二硫键异构酶（PDI）和热休克蛋白70（HSP 70）。最后，SCZJ 3感染鸡胚肾（CEK）细胞被发现抑制抗HSP 70，但不抗PDI的多克隆抗体。这些结果表明，HSP 70是IBV受体复合物的一部分，可能有助于理解S介导的IBV细胞侵入机制。
Avian infectious bronchitis virus (IBV) is a member of the familyCoronaviridae. A binding domain that mediates the attachment of the virus to its receptor has been identified in the S1 protein of prototype IBV strain M41. In this study, we identified this binding domain in a different strain, as well as the cellular proteins that interact with it. First, we expressed the S1Nproteins (residues 19-270) of M41 and another isolate, SCZJ3, and compared the binding capacities of recombinant S1N-M41 and S1N-SCZJ3 to host tissues. Protein histochemistry showed that both S1N-M41 and S1N-SCZJ3 could bind to lung and kidney, and that recombinant S1N-SCZJ3 displayed a distinctive staining pattern in the proventriculus. Recombinant S1N-SCZJ3 was then employed to purify binding-associated proteins in lung, kidney, and proventriculus. Using an affinity chromatography assay, two common bands of about 60 kDa and 70 kDa were obtained from the total tissue proteins. These protein bands were identified by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) as protein disulfide isomerase (PDI) and heat shock protein 70 (HSP70). Finally, infection of chicken embryo kidney (CEK) cells by SCZJ3 was found to be inhibited by anti-HSP70 but not anti-PDI polyclonal antibody. These data indicate that HSP70 is part of the receptor complex of IBV and might help to understand the mechanism of S-mediated cell entry of IBV.
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