Protein Palmitoylation in Yeast and Mammals

酵母和哺乳动物中的蛋白质棕榈酰化

• 批准号: 7367808
• 负责人: NICHOLAS G DAVIS
• 金额: $ 28.36万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-06-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7367808
• 关键词: Address; Amino Acid Permease; Boxing; Brain; Caveolae; Cellular biology; Chitin; Complex; Demyelinating Diseases; Development; Embryo; Endoplasmic Reticulum; Enzymatic Biochemistry; Enzymes; Eukaryotic Cell; Excision; Family; Future; Human Genome; Huntington Disease; Inherited; Integral Membrane Protein; Investigation; Knockout Mice; Ligase; Lipids; Malignant Neoplasms; Mammals; Maps; Membrane; Membrane Microdomains; Membrane Protein Traffic; Methodology; Modification; Monitor; Mus; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Palmitic Acylation Site; Pathogenesis; Pelizaeus-Merzbacher Disease; Play; Process; Protein Family; Proteins; Proteome; Proteomics; Quality Control; Rattus; Regulation; Research; Role; Saccharomyces cerevisiae; Signal Transduction; Stimulus; Substrate Specificity; Surveys; Synapses; Synaptic Transmission; Technology; Transferase; Work; Yeasts; comparative; enzyme substrate; in vivo; insight; interest; neurotransmission; new technology; palmitoylation; tool; trafficking; yeast protein

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.

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，其允许蛋白质与膜的调节性束缚，并且细胞信号传导、癌症、神经元传递和膜运输的许多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广泛的认可，但令人惊讶的是，它仍然没有得到充分的研究。棕榈酰化很难检测和预测。此外，直到最近，棕榈酰加成的酶学仍然是一个完全的黑匣子。前两种棕榈酰化酶，即蛋白质酰基转移酶（PAT），仅在四年前通过酵母中的工作确定。这两个鉴定的酵母PAT共同指向DHHC蛋白家族作为可能的PAT家族。酵母有七种DHHC蛋白，而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蛋白。这种方法首先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相匹配。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节以及神经元的发育和可塑性中起着重要作用。像酵母分析一样，这些第一批哺乳动物分析可能会突出棕榈酰化在大脑中的意想不到的新作用，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HIP 14的神经元棕榈酰化底物，HIP 14是一种DHHC PAT，被认为参与了亨廷顿病。最后一个项目将建立在最近发现的作用棕榈酰化的质量控制机制，在内质网（ER）。特别感兴趣的将是棕榈酰化在质量控制机制中的作用的调查，该机制是脱髓鞘、神经退行性Pelizaeus- Merzbacher病的发病机制的基础。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。