Proteomic Approaches to Protein Fatty Acylation

蛋白质脂肪酰化的蛋白质组学方法

基本信息

批准号：
7910658
负责人：
Julian P Whitelegge
金额：
$ 28.59万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-15 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7910658
关键词：
Address Alzheimer&apos s Disease Biochemistry Biology Biophysics Cell physiology Characteristics Chromatography Code Collection Complement Complex Mixtures Data Detection Development Digestion Dimensions Disease Dissociation Electrons Genotype Goals Human Human Genome Ions Lipids Liquid substance Malignant Neoplasms Mass Spectrum Analysis Measures Membrane Modification Nature Neurodegenerative Disorders Peptides Phase Physical Chemistry Pilot Projects Plasma Plasma Proteins Play Population Post-Translational Protein Processing Property Protein Analysis Protein Chemistry Protein Fragment Proteins Proteolysis Proteome Proteomics Protocols documentation Recovery Relative (related person)Resolution Role Shotguns Site Solubility Technology Time Variant aqueous chemical cleavage experience fatty acylation improved innovation mass spectrometer nano novel protein folding protein protein interaction public health relevance research study reversed phase chromatography tandem mass spectrometry two-dimensional

项目摘要

DESCRIPTION (provided by applicant): Proteomic approaches to protein fatty acylation Protein lipidation is a fundamentally important cellular process that functions in protein folding, protein targeting, membrane interaction and protein/protein interactions. It plays significant roles in cancer and neurodegenerative disease though our understanding of the exact nature of these roles is relatively poor. Discovery of new protein lipidation targets will potentially impact other diseases. Unfortunately, shotgun proteomics workflows that simplify proteins to a collection of peptides tend not to detect lipidated peptides, probably because the protocols employed are optimized to the most easily recovered soluble peptides (proteotypic). Addressing the bias against protein lipidation detection in proteomics requires novel innovations in separations technologies for bottom-up approaches and specialized top-down or middle-down approaches when targeting intact lipidated proteins. Specific aim one will focus on efficient digestion of lipidated proteins and subsequent recovery of lipidated peptides via enrichment with novel HILIC chromatography protocols. We will also innovate modified reverse-phase protocols for recovery of long- chain acylated peptides and tandem mass spectrometry. The overall goal will be a robust two-dimensional separation technology for proteome-wide detection and characterization of lipidated peptides. Specific aim two will focus development of top-down mass spectrometry technologies for characterization of lipidated proteins. This aim includes MSMSMS strategies as well as middle-down strategies to target robust localization of lipidation sites. The overall goal will be a robust mass spectrometry /protein chemistry technology for top-down analysis of protein lipidation across the proteome. PUBLIC HEALTH RELEVANCE: The diversity of proteins coded in the human genome perform the majority of cellular functions that allow human beings to thrive. Some proteins are modified to make them greasy (fatty acylation or lipidation) thereby modulating their function desirably, or in diseases including some cancers and possibly Alzheimer's Disease undesirably. We propose to develop new proteomics technologies to accurately describe and measure this important protein modification that has to date been poorly detected.

描述（由申请人提供）：蛋白质脂肪酰化蛋白脂化的蛋白质组学方法是一种从根本上重要的细胞过程，在蛋白质折叠，蛋白质靶向，膜相互作用和蛋白质/蛋白质相互作用中起作用。尽管我们对这些角色的确切性质的理解相对较差，但它在癌症和神经退行性疾病中起着重要作用。发现新的蛋白质脂化靶标可能会影响其他疾病。不幸的是，将蛋白质简化为一组肽的shot弹枪蛋白质组学工作流往往无法检测到脂质的肽，这可能是因为使用的协议被优化为最容易恢复的可溶性肽（蛋白质型）。解决蛋白质组学中蛋白质脂化检测的偏见需要在靶向完整的脂肪蛋白时，需要在自下而上的方法和专门的自上而下或中向方法的分离技术中进行新的创新。特定目标将专注于有效地消化脂肪蛋白，并通过使用新型的Hilic色谱方案富集来恢复脂质的肽。我们还将创新改进的逆相方案，以恢复长链酰化肽和串联质谱。总体目标将是一种可靠的二维分离技术，用于蛋白质组范围的检测和脂化肽的特征。具体目标两个将集中于自上而下的质谱技术来发展脂肪蛋白的表征。该目的包括MSMSMS策略以及针对脂质位点稳健定位的中东策略。总体目标将是稳健的质谱 /蛋白质化学技术，用于对整个蛋白质组蛋白质脂化的自上而下分析。公共卫生相关性：人类基因组中编码的蛋白质的多样性具有使人类繁衍的大部分细胞功能。一些蛋白质被修饰，使其油腻（脂肪酰化或脂化），从而可以调节其功能，或者不可能地调节包括某些癌症以及可能是阿尔茨海默氏病在内的疾病。我们建议开发新的蛋白质组学技术，以准确描述和测量迄今为止检测到的这种重要的蛋白质修饰。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Mass spectrometric measurements of the apolipoproteins of bovine (Bos taurus) HDL.

牛 (Bos taurus) HDL 载脂蛋白的质谱测量。

DOI：
10.1016/j.cbd.2011.10.001
发表时间：
2012
期刊：
Comparative biochemistry and physiology. Part D, Genomics & proteomics
影响因子：
0
作者：
DellaDonna,Lorenza;Bassilian,Sara;Souda,Puneet;Nebbia,Carlo;Whitelegge,JulianP;Puppione,DonaldL
通讯作者：
Puppione,DonaldL

Proteogenomic Review of the Changes in Primate apoC-I during Evolution.

灵长类 apoC-I 进化过程中变化的蛋白质组学综述。

DOI：
10.1007/s11515-013-1278-7
发表时间：
2013
期刊：
Frontiers in biology
影响因子：
0
作者：
Puppione,Donald;Whitelegge,JulianP
通讯作者：
Whitelegge,JulianP

Mass spectral measurements of the apoHDL in horse (Equus caballus) cerebrospinal fluid.

马 (Equus caballus) 脑脊液中 apoHDL 的质谱测量。

DOI：
10.1016/j.cbd.2012.02.002
发表时间：
2012
期刊：
Comparative biochemistry and physiology. Part D, Genomics & proteomics
影响因子：
0
作者：
Puppione,DonaldL;DellaDonna,Lorenza;Bassilian,Sara;Souda,Puneet;MacDonald,MelindaH;Whitelegge,JulianP
通讯作者：
Whitelegge,JulianP

Structural biology. Up close with membrane lipid-protein complexes.

结构生物学。