Chemical mapping of the PPIase interactome in C. elegans - development of a systems biology toolbox

秀丽隐杆线虫（秀丽隐杆线虫）是一种由959个细胞组成的自由活的圆形蠕虫。它的基因组产生19000种蛋白质（相比之下，人类的基因组大约有25000种）。我们将把化学物质引入蠕虫体内，观察它们的反应，尤其是它们蛋白质的变化。小分子可能是激素、药物或毒素，对生物系统有深远的影响。变化通常是由小分子（配体）直接与特定的“目标蛋白质”结合并阻止其发挥正常功能引起的。这些类型的分子被称为抑制剂。目前，当一个给定的蛋白质与配体结合时，我们对细胞中的“连锁”效应了解甚少。我们想设计一种分子，可以结合秀丽隐杆线虫的特定蛋白质家族，然后改变它们的行为。我们将监测化学品对整个蠕虫的影响，并确定哪些其他蛋白质受到影响。在这个项目中，我们将只研究一个蛋白质家族，即亲免疫蛋白，对此我们有相当多的背景知识。超过20种不同的蛋白质属于这个家族。我们知道它们可以起到酶的作用，也可以帮助蛋白质折叠。我们已经合成了抑制这些亲免疫蛋白的新分子，并在秀丽隐杆线虫中显示出生物活性。我们已经在一些抑制剂分子上贴上了荧光标记，这样我们就可以（用光学显微镜）看到这些分子被蠕虫吸收的地方。我们将看到蠕虫体内的蛋白质在暴露于这些抑制剂配体时是如何变化的。通过测试数百种不同的抑制剂，我们将在这个项目中确定具有特定生物效应的分子。秀丽隐杆线虫中的许多蛋白质与哺乳动物中的相似，因此我们也将了解这些化学物质对动物系统的影响。我们从蠕虫身上获得的数据可能对人类和兽医生物学在基因调控、发育生物学和药物发现等领域产生重大影响。

Caenorhabditis elegans (C. elegans) is a free living round worm comprising only 959 cells. Its genome produces 19,000 proteins (compared with some 25,000 from the human genome). We will introduce chemicals into the worms and look to see how they respond, particularly in terms of changes in their proteins. Small molecules which might be hormones, drugs or toxins can have profound effects on biological systems. Changes are usually caused by the small molecules (ligands) binding directly to a particular 'target protein' and stopping it from performing its normal function. These types of molecules are known as inhibitors. Currently we have a very poor understanding of the 'knock-on' effects in the cell when a given protein is bound to a ligand. We want to design molecules that bind to particular families of proteins in C.elegans and then modify their behaviour. We will monitor the effect of the chemical on the whole worm and identify which of the other proteins are affected. In this project we will only look at one family of proteins, the immunophilins, for which we have considerable background knowledge. Over 20 different proteins belong to this family. We know that they can act as enzymes and also help protein folding. We have synthesised new classes of molecules that inhibit these immunophilins and also show biological activity in the C.elegans. We have already put fluorescent-labels on some of these inhibitor molecules so that we see (using an optical microscope) where the molecules are taken up by the worm. We will see how proteins in worm change when they are exposed to these inhibitor ligands. By testing the many hundreds of different inhibitors that will be made in this project, we will identify molecules that have specific biological effects. Many of the proteins in C. elegans are similar to those in mammals, and so we will also learn about the effect of these chemicals in animal systems. Our data from worms could have a major impact on human and veterinary biology in areas like gene regulation, developmental biology, and drug discovery.