STRUCTURAL ANALYSIS OF SPRINTER

SPRINTER的结构分析

• 批准号: 7609824
• 负责人: ERICA M SELVA
• 金额: $ 6.59万
• 依托单位: UNIVERSITY OF DELAWARE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7609824
• 关键词: Amino Acids; Cells; Collaborations; Complex; Computer Retrieval of Information on Scientific Projects Database; Delaware; Drosophila genus; Funding; Grant; Institution; Integral Membrane Protein; Laboratories; Lead; Ligands; N-terminal; Names; Nuclear Magnetic Resonance; Process; Protein Region; Proteins; Research; Research Personnel; Resources; Source; Structure; Transmembrane Domain; United States National Institutes of Health; Universities; X-Ray Crystallography; extracellular; in vivo; novel; tool

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. Recently, we have identified a novel multi-transmembrane protein in Drosophila we named Sprinter (Srt), which is required for the secretion of active Wg/Wnt from ligand expressing cells. Although it is known that Srt is necessary for secretion of active Wg/Wnt, its actual function in the complex maturation process of Wnts is not yet understood. Analysis of the Srt amino acid suggests it could have several possible topologies. We hypothesize Srt has 4 transmembrane domains with a large N-terminal globular extracellular/luminal domain which likely constitutes the predominate functional region of this protein. This region includes a conserved structural domain of unknown function, DUFF1141. The overall objective of this proposal is to initiate structural and functional characterization of the Srt globular extracellular/luminal domain and determine the overall topology of Srt. To do this we will first perform topological studies to confirm our predicted structure. Next, we will develop constructs that will allow for high levels of expression of N-terminal Srt for its purification, as well as its in vivo expression. We will use these tools in order to ascertain the function of the N-terminal region in vivo and determine if Srt and its N-terminal region interact with Wg/Wnt and possibly other proteins involved in Wnt processing. Ultimately we believe these studies will lead to collaboration with a laboratory here at the University of Delaware to perform structural analysis of the purified Srt N-terminal globular domain by X-ray crystallography or nuclear magnetic resonance to the integrate structural and functional aspects of Srt.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 最近，我们在果蝇中发现了一种新型的多跨膜蛋白，我们将其命名为 Sprinter (Srt)，它是配体表达细胞分泌活性 Wg/Wnt 所必需的。尽管已知 Srt 对于活性 Wg/Wnt 的分泌是必需的，但其在 Wnts 复杂成熟过程中的实际功能尚不清楚。对 Srt 氨基酸的分析表明它可能有几种可能的拓扑结构。我们假设 Srt 有 4 个跨膜结构域，其中有一个大的 N 端球状细胞外/管腔结构域，这可能构成了该蛋白的主要功能区域。该区域包括一个功能未知的保守结构域 DUFF1141。该提案的总体目标是启动 Srt 球状细胞外/腔域的结构和功能表征，并确定 Srt 的整体拓扑。为此，我们将首先进行拓扑研究以确认我们预测的结构。接下来，我们将开发能够高水平表达 N 端 Srt 的构建体，以进行纯化以及体内表达。我们将使用这些工具来确定体内 N 末端区域的功能，并确定 Srt 及其 N 末端区域是否与 Wg/Wnt 以及可能参与 Wnt 加工的其他蛋白质相互作用。最终，我们相信这些研究将导致与特拉华大学实验室的合作，通过 X 射线晶体学或核磁共振对纯化的 Srt N 末端球状结构域进行结构分析，以整合 Srt 的结构和功能方面。