Influence of Palmitoylation on GSTP1 Structure, Function, and Cellular Localization.

棕榈酰化对 GSTP1 结构、功能和细胞定位的影响。

• 批准号: RGPIN-2017-06154
• 负责人: Leslie, Elaine
• 金额: $ 2.04万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=682459
• 关键词: Influence; Palmitoylation; GSTP1; Structure; Function

Glutathione transferases (GSTs) are a superfamily of proteins best known for detoxifying harmful electrophilic compounds by catalyzing their conjugation with glutathione. GSTP1 is one member of this superfamily. In addition to an important role in detoxification, GSTP1 is involved in the regulation of cell signalling. The location of a protein such as GSTP1 within a cell can have a significant effect on how the protein functions. Cells are organized into different compartments suspended in a cellular fluid, all of which are contained within the outer skin known as the plasma membrane. Studies previously suggested that GSTP1 is located in the cellular fluid, but we have discovered that GSTP1 is also located in cellular membranes. We have recently discovered that GSTP1 can be modified with a fatty molecule called palmitate. Palmitate modification is likely responsible for moving the location of GSTP1 from the cellular fluid to cell membranes and might have additional functions. We are trying to determine precisely where palmitoylation is occurring on GSTP1 so that we can generate a nonpalmitate modified GSTP1 mutant. We are trying to understand the influence the palmitate modification has on the structure of GSTP1 and we are using multiple biochemical approaches to investigate this. In addition, the influence of palmitate on the catalytic and non-catalytic functions of GSTP1 will be investigated. This will include the investigation of how GSTP1 tagged with palmitate interacts differently with other proteins. We will also compare how the palmitate modified GSTP1 interacts with pure lipids and membranes isolated from cell lines versus the nonmodified GSTP1. Lastly, we will determine how palmitate tagging influences where GSTP1 is located in the cell. Knowledge gained from this research will lead to novel insights into the fundamental biology of GSTP1 and how palmitate modification influences its structure, function, membrane association, and cellular localization. The GSTP1 form found in cellular fluid is well characterized, however, virtually nothing is known about the palmitate-modified form. This research will provide novel information for the diverse functions of GSTP1 in cellular detoxification and signalling. In addition, results from these studies will have far reaching implications for understanding the influence of palmitate modification on the structure, function, and membrane association of other proteins.

谷胱甘肽转移酶(GSTs)是一个蛋白质超家族，最著名的功能是通过催化它们与谷胱甘肽的结合来解毒有害的亲电化合物。GSTP1是这个超家族的成员之一。除了在解毒方面发挥重要作用外，GSTP1还参与细胞信号的调节。GSTP1等蛋白质在细胞内的位置对蛋白质的功能有很大影响。细胞被组织成悬浮在细胞液中的不同隔室，所有这些隔室都包含在被称为质膜的外层皮肤内。先前的研究表明GSTP1位于细胞液中，但我们发现GSTP1也位于细胞膜中。我们最近发现，GSTP1可以被一种名为棕榈酸酯的脂肪分子修饰。棕榈酸酯修饰可能负责将GSTP1的位置从细胞液移动到细胞膜上，并可能具有额外的功能。我们正试图准确地确定GSTP1上发生棕榈酰化的位置，以便我们能够产生非棕榈酸酯修饰的GSTP1突变体。我们正试图了解棕榈酸酯修饰对GSTP1结构的影响，我们正在使用多种生化方法来研究这一点。此外，还将研究棕榈酸酯对GSTP1催化和非催化功能的影响。这将包括研究用棕榈酸酯标记的GSTP1如何与其他蛋白质以不同方式相互作用。我们还将比较棕榈酸酯修饰的GSTP1与未修饰的GSTP1与从细胞系分离的纯脂和膜相互作用的方式。最后，我们将确定棕榈酸标记如何影响GSTP1在细胞中的位置。从这项研究中获得的知识将导致对GSTP1的基础生物学以及棕榈酸酯修饰如何影响其结构、功能、膜结合和细胞定位的新见解。在细胞液中发现的GSTP1形式具有很好的特性，然而，关于棕榈酸酯修饰的形式几乎一无所知。本研究将为GSTP1在细胞解毒和信号转导中的不同功能提供新的信息。此外，这些研究的结果将对理解棕榈酸酯修饰对其他蛋白质的结构、功能和膜结合的影响具有深远的意义。