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A chemical genetic approach to the analysis of peroxisome biogenesis

分析过氧化物酶体生物发生的化学遗传学方法

基本信息

批准号：
BB/E013740/1
负责人：
Alison Baker
金额：
$ 69.82万
依托单位：
University of Leeds
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2007
资助国家：
英国
起止时间：
2007 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FE013740%2F1
关键词：
chemical genetic approach analysis peroxisome

项目摘要

All organisms are made up of cells. Complex organisms like plants and animals have many different types of cells organised into tissues and organs that perform different functions. Examples are a plant leaf, that carries out photosynthesis and the human heart that pumps blood around our body. Each of these and many other different processes require not only the interaction between cells but the proper organisation of chemical reactions within cells. Cells of all but the simplest organisms contain compartments called organelles. These compartments require proteins in order to function. Proteins made within the cytoplasm of the cell are delivered to organelles by protein trafficking pathways. The proteins carry signals that are recognised by other proteins that sort and deliver them to the correct place. One of these organelles is the peroxisome. If peroxisomes go wrong the consequences are dire. Children born with defective peroxisomes suffer very serious disabilities and usually die. Plants with completely defective peroxisomes cannot produce viable seeds. Because of the importance of protein trafficking pathways, scientists want to understand how they work in detail. We already know a lot, mainly from knocking out the function of genes that make proteins that are needed for these pathways. But sometimes knocking out genes causes the organism to die, which isn't very useful, or has no effect, because there may be other genes that can perform the same function as the defective gene. Small chemicals that can enter cells and inhibit the function of some of these proteins can offer a very useful alternative way to understand how these processes work, because chemicals can be added at specific time points and taken away later on. They can be added in different amounts so that they do not have a lethal effect and the effects can be monitored from the moment the chemical is added. If there is more than one similar protein carrying out the same function the same chemical will probably affect all the similar proteins, whereas knocking out a gene will only affect the protein made by that one gene. Nature, augmented by synthetic chemistry, has produced literally millions of chemical structures. In theory there should be a small chemical that interferes with the function of every protein in every cell. We have identified a small group of chemicals which interfere with the import of proteins into peroxisomes and another group which alter how peroxisomes move around the cell. A further compound blocks the breakdown of fats by the peroxisome. The aim of this project is identify which specific parts of the cellular machinery these molecules are interacting with, so that we can understand more about the precise details of how the mechanism and regulation of proteins import into peroxisomes. Firstly we will understand the precise molecular features of the small molecules that are necessary to cause the observed effect. At the same time we will characterise the effects of each molecule in detail, for example working out how much molecule is needed to cause each type of response and also if exposure to the chemical changes the levels of certain proteins known to be involved in peroxisomal protein trafficking. We will then use a range of different chemical and biochemical techniques to identify the precise biomolecular target of the small molecules. By understanding exactly which biomolecule the small molecule binds to we will gain new insights into the operation of the cell. Understanding these processes better could eventually lead to the development of new drugs or the ability to manipulate these pathways in plants or micro-organisms for biotechnological purposes.

所有的有机体都是由细胞组成的。像植物和动物这样的复杂生物体有许多不同类型的细胞，它们被组织成执行不同功能的组织和器官。例如，进行光合作用的植物叶子，以及将血液输送到我们身体周围的人类心脏。这些过程中的每一个以及许多其他不同的过程不仅需要细胞之间的相互作用，还需要细胞内化学反应的适当组织。除最简单的有机体外，所有生物的细胞都含有称为细胞器的隔间。这些隔室需要蛋白质才能发挥作用。在细胞质中产生的蛋白质通过蛋白质运输途径被运送到细胞器。这些蛋白质携带被其他蛋白质识别的信号，这些蛋白质将它们分类并传递到正确的位置。这些细胞器中的一个是过氧化物体。如果过氧化物酶出现问题，后果将是可怕的。出生时有过氧化物酶缺陷的儿童会遭受非常严重的残疾，通常会死亡。具有完全缺陷的过氧化物体的植物不能产生可存活的种子。由于蛋白质运输途径的重要性，科学家们想要详细了解它们是如何工作的。我们已经知道了很多，主要是通过敲除基因的功能，这些基因产生了这些途径所需的蛋白质。但有时敲除基因会导致有机体死亡，这并不是很有用，或者没有作用，因为可能有其他基因可以执行与缺陷基因相同的功能。可以进入细胞并抑制其中一些蛋白质功能的小化学物质可以提供一个非常有用的替代方法来了解这些过程是如何工作的，因为化学物质可以在特定的时间点添加，然后再带走。它们可以按不同的量添加，这样它们就不会产生致命的影响，并且可以从添加化学物质的那一刻起就对其影响进行监测。如果有不止一种相似的蛋白质具有相同的功能，相同的化学物质可能会影响所有相似的蛋白质，而敲除一个基因只会影响由该基因制造的蛋白质。大自然，加上合成化学，已经产生了数以百万计的化学结构。从理论上讲，应该有一种小的化学物质干扰每个细胞中每种蛋白质的功能。我们已经确定了一小群化学物质，它们干扰蛋白质输入到过氧化物体中，另一组化学物质改变了过氧化物体在细胞内移动的方式。另一种化合物阻止了过氧化物体对脂肪的分解。这个项目的目的是确定这些分子与细胞机械的哪些特定部分相互作用，以便我们能够更多地了解蛋白质如何输入到过氧化酶体的机制和调控的确切细节。首先，我们将了解引起观察到的效应所必需的小分子的精确分子特征。与此同时，我们将详细描述每个分子的影响，例如计算出需要多少分子才能引起每种类型的反应，以及如果暴露在化学物质中会改变某些已知参与过氧化体蛋白运输的蛋白质的水平。然后，我们将使用一系列不同的化学和生化技术来识别小分子的精确生物分子靶标。通过准确地了解小分子结合到哪个生物分子上，我们将获得对细胞运行的新见解。更好地了解这些过程最终可能导致新药的开发，或者为生物技术目的在植物或微生物中操纵这些途径的能力。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The life of the peroxisome: from birth to death.

过氧化物酶体的生命：从出生到死亡。

DOI：
10.1016/j.pbi.2014.09.003
发表时间：
2014
期刊：
Current opinion in plant biology
影响因子：
9.5
作者：
Baker A
通讯作者：
Baker A

The Breakdown of Stored Triacylglycerols Is Required during Light-Induced Stomatal Opening.

DOI：
10.1016/j.cub.2016.01.019
发表时间：
2016-03-07
期刊：
Current biology : CB
影响因子：
0
作者：
McLachlan DH;Lan J;Geilfus CM;Dodd AN;Larson T;Baker A;Hõrak H;Kollist H;He Z;Graham I;Mickelbart MV;Hetherington AM
通讯作者：
Hetherington AM

PEX14 binding to Arabidopsis PEX5 has differential effects on PTS1 and PTS2 cargo occupancy of the receptor.