Functional analysis of a family of TANGO1-like proteins. What is the influence on transport of large cargos and are so-called megacarriers generated?

TANGO1 样蛋白家族的功能分析。

• 批准号: 201191293
• 负责人: Dr. Patrik Erlmann
• 金额: --
• 依托单位: Center for Genomic Regulation
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2011
• 资助国家: 德国
• 起止时间: 2010-12-31 至 2012-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/201191293?language=en
• 关键词: Functional; analysis; family; TANGO1; like

Higher eukaryotes produce a wide range of proteins and molecules, a large portion of these have their destination outside of the cell. In order to reach the outside, these proteins are translated into the endoplasmatic reticulum, are transported to the Golgi and from there to the cell surface. The transport from the endoplasmatic reticulum to the Golgi is facilitated by COPII vesicles, that have a defined size of about 60-90nm. Interestingly, some of the transported proteins are quit large. An example is Collagen VII that comes in the form of a 400nm long, rigid triple helix. It has been speculated that these large cargos are transported in megacarriers, larger than usual vesicles. So far it is neither known whether these vesicles truly exist, nor how they form.It is known however, that the ER exit site localized protein TANGO1 plays a role in the transport of Collagen VII. When TANGO1 is depleted, cells can no longer shuttle Collagen VII from the endoplasmatic reticulum to the Golgi. Interestingly, bulk protein secretion is not affected in these cells. Trough sequence comparison I discovered a family of TANGO1-like (Tali) proteins that like TANGO1 localize to ER exit sites. I want to explore if these proteins, together with TANGO1, play a role in the generation of megacarriers or whether they are needed in other steps of vesicle biogenesis at the endoplasmatic reticulum.An additional aim of the proposal is to investigate how large proteins are transported anyway. To clarify the involvement of megacarriers in the transport of large cargo and to gain insight into the mechanism, I will in Vitro reconstitute vesicle biogenesis at the level of the ER.

高等真核生物产生广泛的蛋白质和分子，其中很大一部分的目的地是细胞外。为了到达外部，这些蛋白质被翻译到内质网，被运送到高尔基体，并从那里到达细胞表面。从内质网到高尔基体的运输由COPII囊泡促进，其具有约60- 90 nm的确定尺寸。有趣的是，一些转运的蛋白质非常大。一个例子是VII型胶原蛋白，它以400 nm长的刚性三重螺旋的形式存在。据推测，这些大的货物是在megacarriers，比通常的囊泡更大的运输。到目前为止，这些囊泡是否真的存在，也不知道它们是如何形成的。然而，已知ER出口位点定位的蛋白质TANGO 1在胶原蛋白VII的运输中起作用。当TANGO 1耗尽时，细胞不能再将胶原VII从内质网运送到高尔基体。有趣的是，在这些细胞中，大量蛋白质分泌不受影响。通过序列比较，我发现了一个TANGO 1样（Tali）蛋白家族，该家族与TANGO 1一样定位于ER出口位点。我想探索这些蛋白质，与TANGO 1一起，是否在巨载体的产生中发挥作用，或者它们是否在内质网的囊泡生物发生的其他步骤中需要。该提案的另一个目的是调查无论如何大蛋白质是如何运输的。为了阐明巨载体参与运输大型货物，并深入了解其机制，我将在体外重组囊泡生物发生在ER的水平。