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Torreyanic acid: Affinity chromatographic identification of receptors and biochemical analysis of the torreyanic acid-eIF-4A complex .

机译:Torreyanic acid:受体的亲和色谱鉴定和toryanyanic acid-eIF-4A复合物的生化分析。

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摘要

Affinity chromatography was used to identify binding proteins for two natural products: brefeldin A and torreyanic acid. Evidence is presented that brefeldin A binds to two proteins not related to its classic biological activity in protein secretion: crm1/xpoI and L-type calcium channel α 2-subunit. The possible consequences of these interactions are disscussed. Several proteins are identified based on specific binding to a torreyanic acid affinity reagent. The eukaryotic translation initiation factor 4A (eIF-4A) was identified as a probable effector molecule of torreyanic acid action. (eIF-4A) is an RNA-dependent helicase from the DEAD box protein family. It is required for cap-dependent and cap-independent translation. Evidence for a covalent torreyanic acid-eIF-4A adduct is presented. Biochemical analysis of torreyanic acid-eIF-4A complex was performed in vitro and in vivo. Torreyanic acid was shown to inhibit the ATPase, ATP-binding and RNA-binding activities of eIF-4A. A consequence of this interaction is that under specific conditions, torreyanic acid both inhibits and stimulates translation in vivo and in vitro. A model for this dualism is presented. Torreyanic acid is the first identified endogenous ligand for eIF-4A and small molecule effector or translation initiation. Torreyanamide, the bis-diethylamide of torreyanic acid, was synthesized. It has the same biological profile as torreyanic acid, but is more potent.
机译:亲和色谱法用于鉴定两种天然产物的结合蛋白:布雷菲德菌素A和Torryanic酸。有证据表明,布雷菲德菌素A结合两种与其蛋白分泌中的经典生物学活性无关的蛋白:crm1 / xpoI和L型钙通道α2-亚基。讨论了这些相互作用的可能后果。基于与对苯二酸亲和试剂的特异性结合,鉴定了几种蛋白质。真核翻译起始因子4A(eIF-4A)被确定为可能的toryanyanic酸效应分子。 (eIF-4A)是DEAD盒蛋白家族的一种RNA依赖性解旋酶。依赖于上限和依赖于上限的翻译是必需的。提供了共价花胶酸-eIF-4A加合物的证据。在体外和体内进行了对甲苯二酸-eIF-4A复合物的生化分析。示出花梨酸抑制eIF-4A的ATP酶,ATP结合和RNA结合活性。这种相互作用的结果是,在特定条件下,鸟梨酸在体内和体外均抑制和刺激翻译。提出了这种二元论的模型。 Torreyanic Acid是最早鉴定出的eIF-4A和小分子效应子或翻译起始的内源性配体。合成了鸟眼强酸的双二乙酰胺Torreyanamide。它具有与鸟梨酸相同的生物学特性,但更有效。

著录项

  • 作者

    Justman, Craig Jerome.;

  • 作者单位

    Harvard University.;

  • 授予单位 Harvard University.;
  • 学科 Chemistry Biochemistry.
  • 学位 Ph.D.
  • 年度 2000
  • 页码 140 p.
  • 总页数 140
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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