Protein Palmitoylation in Yeast and Mammals

描述（由申请人提供）：蛋白质棕榈酰化是一种可逆的翻译后脂质修饰，可调节蛋白质与膜的拴系，许多细胞信号传导、癌症、神经元传递和膜运输的关键蛋白质被棕榈酰化。尽管棕榈酰化的重要性得到了公认，但令人惊讶的是，它的研究仍然不足。棕榈酰化很难检测和预测。此外，直到最近，棕榈酰添加的酶学仍然是一个完全的黑盒子。前两种棕榈酰化酶，即蛋白酰基转移酶（PATs），仅在四年前在酵母中被发现。这两个鉴定的酵母PAT共同指向了DHHC蛋白家族作为一个可能的PAT家族。酵母中有7种DHHC蛋白，而人类基因组中有23种可识别。为了促进棕榈酰化的全球方法，我们开发了一种新的蛋白质组学方法，从复杂的蛋白质提取物中纯化和鉴定棕榈酰蛋白。该方法首次在酵母中用于鉴定许多新的棕榈酰蛋白，并将各种棕榈酰蛋白与其同源修饰PAT进行匹配。一个新的项目将应用相同的蛋白质组学技术来理解哺乳动物大脑中的棕榈酰化。棕榈酰化在突触信号的调控以及神经元的发育和可塑性中都起着重要的作用。就像酵母分析一样，这些首次对哺乳动物的分析可能会突出棕榈酰化在大脑中意想不到的新作用，为未来的神经科学研究提供刺激。此外，该技术还将用于鉴定HIP14的神经元棕榈酰化底物，HIP14是一种DHHC PAT，已被认为参与亨廷顿病。最后一个项目将建立在最近发现的棕榈酰化在内质网（ER）的质量控制机制中的作用。特别感兴趣的是棕榈酰化在脱髓鞘、神经退行性Pelizaeus- Merzbacher病发病机制的质量控制机制中的作用。因此，除了扩大对这种脂质修饰的细胞生物学的基本理解外，我们还希望深入了解棕榈酰化在神经退行性疾病中的作用。

DESCRIPTION (provided by applicant): Protein palmitoylation is a reversible post-translational lipid modification that allows regulated tethering of proteins to membranes and many of key proteins of cellular signaling, cancer, neuronal transmission, and membrane trafficking are palmitoylated. Despite its well-recognized importance, palmitoylation remains surprisingly understudied. Palmitoylation is difficult to detect and predict. Furthermore, until quite recently, the enzymology of palmitoyl addition had remained a total black box. The first two palmitoylation enzymes, i.e. protein acyl transferases (PATs), were identified only four years ago by work in yeast. The two identified yeast PATs together pointed towards the DHHC protein family as a possible PAT family. Yeast has seven of these DHHC proteins, while 23 are identifiable from the human genome. To facilitate global approaches to palmitoylation, we have developed a new proteomic methodology that purifies and identifies palmitoyl- proteins from complex protein extracts. This approach has first been used in yeast to identify many new palmitoyl-proteins and also to match the various palmitoyl-proteins with their cognate modifying PAT. A new project will apply the same proteomic technologies towards an understanding of palmitoylation in the mammalian brain. Palmitoylation plays a prominent role both in the regulation of synaptic signaling as well as in neuronal development and plasticity. Like the yeast analyses, these first mammalian analyses likely will highlight unanticipated new roles for palmitoylation in the brain, providing stimulus to future neuroscientific research. In addition, this technology also will be used to identify the neuronal palmitoylation substrates of HIP14, a DHHC PAT that has been implicated as participating in Huntington's disease. A final project will build on a recently discovered role for palmitoylation in the quality control mechanisms that act at the endoplasmic reticulum (ER). Of particular interest will be an investigation of the role of palmitoylation in the quality control mechanisms that underlie pathogenesis of demyelinating, neurodegenerative Pelizaeus- Merzbacher disease. Thus, in addition to expanding basic understanding of the cell biology of this lipid modification, we also hope to gain insight into the role of palmitoylation in neurodegenerative disease.