Protein Palmitoylation in Yeast and Mammals

描述(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，允许蛋白质受控地与膜连接，许多细胞信号、癌症、神经元传递和膜运输的关键蛋白质都是棕榈酰化的。尽管棕榈酰化的重要性得到了公认，但令人惊讶的是，对它的研究仍然不足。棕榈酰化很难检测和预测。此外，直到最近，棕榈酰加成的酶学一直是一个完全的黑匣子。最初的两种棕榈酰化酶，即蛋白质酰基转移酶(PATS)，仅在四年前通过在酵母中的工作而被鉴定出来。这两个鉴定的酵母片一起指向DHHC蛋白家族，可能是PAT家族。酵母中有7种这种DHHC蛋白，其中23种可以从人类基因组中识别出来。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化蛋白质。这种方法已首次用于酵母中，以鉴定许多新的棕榈酰基蛋白质，并将各种棕榈酰基蛋白质与它们的同源修饰PAT进行匹配。一个新的项目将应用同样的蛋白质组学技术来理解哺乳动物大脑中的棕榈酸化。棕榈酰化在突触信号的调节以及神经元的发育和可塑性中都起着重要的作用。像酵母菌分析一样，这些第一批哺乳动物分析可能会突出棕榈酸化在大脑中出人意料的新作用，为未来的神经科学研究提供刺激。此外，这项技术还将用于鉴定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)质量控制机制中的作用基础上。尤其令人感兴趣的是研究棕榈酰化在脱髓鞘、神经退行性Pelizaeus-Merzbacher病发病机制中的质量控制机制的作用。因此，除了扩大对这种脂质修饰的细胞生物学的基本了解外，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.