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首页> 外文期刊>Biochemistry >Direct Interaction of the Mouse Cytomegalovirus m152/gp40 Immunoevasin with RAE-1 Isoforms
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Direct Interaction of the Mouse Cytomegalovirus m152/gp40 Immunoevasin with RAE-1 Isoforms

机译：小鼠巨细胞病毒m152 / gp40免疫血管素与RAE-1亚型的直接相互作用

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Cytomegaloviruses (CMVs) are ubiquitous species-specific viruses that establish acute, persistent,nand latent infections. Both human and mouse CMVs encode proteins that inhibit the activation of naturalnkiller (NK) cells by downregulating cellular ligands for theNKcell activating receptor,NKG2D. TheMCMVnglycoprotein m152/gp40 downregulates the surface expression of RAE-1 to prevent NK cell control in vivo.nSo far, it is unclear if there is a direct interaction between m152 and RAE-1 and, if so, if m152 interactsndifferentially with the five identified RAE-1 isoforms, which are expressed as two groups in MCMV-nsusceptible or -resistantmouse strains. To address these questions, we expressed and purified the extracellularndomains of RAE-1 and m152 and performed size exclusion chromatography binding assays as well asnanalytical ultracentrifugation and isothermal titration calorimetry to characterize these interactions quanti-ntatively. We further evaluated the role of full-length and naturally glycosylated m152 and RAE-1 inncotransfected HEK293T cells. Our results confirmed that m152 binds RAE-1 directly, relatively tightlyn(Kd<5 μM), and with 1:1 stoichiometry. The binding is quantitatively different depending on particularnRAE-1 isoforms, corresponding to the susceptibility to downregulation by m152. A PLWY motif found innRAE-1β, although contributing to its affinity form152, does not influence the affinity ofRAE-1γ orRAE-1δ,nsuggesting that other differences contribute to the RAE-1-m152 interaction. Molecular modeling of thendifferent RAE-1 isoforms suggests a potential site for the m152 interaction.
机译：巨细胞病毒（CMV）是普遍存在的物种特异性病毒，可引起急性，持续性和潜伏性感染。人类和小鼠CMV都编码通过下调NK细胞活化受体NKG2D的细胞配体来抑制自然NK（NK）细胞活化的蛋白质。 MCMV糖蛋白m152 / gp40下调RAE-1的表面表达以防止体内NK细胞的控制。 RAE-1亚型，在MCMV易感性或抗性小鼠品系中表示为两组。为了解决这些问题，我们表达并纯化了RAE-1和m152的胞外域，并进行了体积排阻色谱结合测定以及超离心分析和等温滴定量热法，以定量表征这些相互作用。我们进一步评估了全长和天然糖基化的m152和RAE-1 inncotransfected HEK293T细胞的作用。我们的结果证实，m152与RAE-1直接结合，相对紧密（Kd <5μM），化学计量比为1：1。结合在数量上有所不同，具体取决于特定的nRAE-1亚型，对应于m152对下调的敏感性。在nRAE-1β中发现的PLWY基序虽然有助于其亲和形式152，但不影响RAE-1γ或RAE-1δ的亲和性，暗示其他差异也有助于RAE-1-m152相互作用。随后不同的RAE-1亚型的分子模型表明了m152相互作用的潜在位点。

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    《Biochemistry》 |2010年第11期|p.2443-2453|共11页
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    Li Zhi Janet Mans Michael J. Paskow Patrick H. Brown Peter Schuck Stipan Jonji Kannan Natarajan and David H. Margulies;

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    ‡Molecular Biology Section, Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, and§Dynamics of Macromolecular Assembly, Laboratory of Bioengineering and Physical Science, National Institute of BiomedicalImaging and Bioengineering, National Institutes of Health, Bethesda, Maryland 20892,) University of the Witwatersrand,Johannesburg 2050, South Africa, and ^Department of Histology and Embryology, Faculty of Medicine, University of Rijeka,51000 Rijeka, Croatia.@Current address: Department of Medical Virology, University of Pretoria, Pretoria 0001, South Africa.;

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