STRUCTURAL ANALYSIS OF EHD1

EHD1的结构分析

• 批准号: 7721677
• 负责人: PAUL L SORGEN
• 金额: $ 0.27万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721677
• 关键词: Amino Acids; Asparagine; Binding; C-terminal; Calcium Binding; Cell membrane; Cells; Charge; Computer Retrieval of Information on Scientific Projects Database; Endosomes; Funding; Grant; Helix (Snails); Institution; Lipids; Localized; Mediating; Membrane; Modeling; N-terminal; Nuclear Magnetic Resonance; Phenylalanine; Phosphatidylinositols; Phospholipids; Positioning Attribute; Proline; Property; Proteins; Recycling; Research; Research Personnel; Resources; Side; Source; Stretching; Tubular formation; United States National Institutes of Health; ear helix; inorganic phosphate; receptor recycling; two-dimensional

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The C-terminal Eps15 homology domain-containing protein, EHD1, regulates the recycling of receptors from the endocytic recycling compartment to the plasma membrane. In cells, EHD1 localizes to tubular and spherical recycling endosomes. To date, the mode by which EHD1 associates with endosomal membranes remains unknown, and it has not been determined whether this interaction is direct or via interacting proteins. Here, we provide evidence demonstrating that EHD1 has the ability to bind directly and preferentially to an array of phospholipids, preferring phosphatidylinositol lipids with a phosphate at position 3. Previous studies have demonstrated that EH-domains coordinate calcium binding and interact with proteins containing the tripeptide asparagine-proline-phenylalanine (NPF). Using two-dimensional Nuclear Magnetic Resonance analysis, we now describe a new function for the Eps15 homology (EH) domain of EHD1 and show that it is capable of directly binding phosphatidylinositol moieties. Moreover, we have expanded our studies to include the C-terminal EH-domain of EHD4 and the second of the three N-terminal EH-domains of Eps15, and have demonstrated that phosphatidylinositol-binding may be a more general property shared by other EH-domains. Our analysis reveals a model by which the side chains of several positively charged residues within a 30 amino acid stretch localized primarily to the second and third helices of the EH-domain mediate a direct interaction with phosphatidylinositols.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 C-末端Eps 15同源结构域的蛋白质，EHD 1，调节受体从内吞再循环室到质膜的再循环。在细胞中，EHD 1定位于管状和球形再循环内体。迄今为止，EHD 1与内体膜结合的模式仍然未知，并且尚未确定这种相互作用是直接的还是通过相互作用的蛋白质。在这里，我们提供的证据表明，EHD 1有能力直接和优先结合的磷脂阵列，首选磷脂酰肌醇脂质与磷酸在位置3。以前的研究表明，EH结构域协调钙结合，并与含有天冬酰胺-脯氨酸-苯丙氨酸（NPF）三肽的蛋白质相互作用。使用二维核磁共振分析，我们现在描述了一个新的功能EHD 1的Eps 15同源（EH）域，并表明它能够直接结合磷脂酰肌醇部分。此外，我们已经扩大了我们的研究，包括C-末端EH-结构域的EHD 4和第二个的三个N-末端EH-结构域的Eps 15，并已证明，磷脂酰肌醇结合可能是一个更普遍的属性共享的其他EH-结构域。我们的分析揭示了一个模型，通过该模型，在一个30个氨基酸的延伸内的几个带正电荷的残基的侧链主要定位于EH-结构域的第二和第三螺旋介导与磷脂酰肌醇的直接相互作用。