Shared Resource Cores

SU MMARY项目(见说明)： 分析糖共轭结构/功能需要来自不同学科的专业知识。 这些工具和方法中的许多不仅困难，而且需要昂贵和复杂的设备。我们已经建立了我们的共享资源核心，不仅包含了对于这次PEG的目标最重要的组成部分，而且还包括建立在我们参与调查人员独特优势基础上的组成部分。每个子核心在其单独的部分中进行了详细说明。 次核心01-质谱学-博士。Van Eyk和Zhang是蛋白质和糖蛋白的质谱分析领域的领导者。核心的装备异常精良。核心产品包括：1)鉴定蛋白质；2)利用固相萃取糖肽对N-连接或O-连接的糖蛋白进行全球定量；3)凝集素亲和/MS分析；4)通过凝集素微阵列监测糖基化的变化；5)使用“Click”化学分析代谢产生的糖结合物；6)通过MALDI-MS和GlycanAnalyser分析糖链；7)细胞表面捕获技术，分离糖蛋白；2)从细胞浆或细胞外环境中定向捕获可溶性糖蛋白；3)MRM/MS分析，有针对性地定量糖蛋白和糖基化位点。 亚核C2-核磁共振--如有可能，糖链结构最好由核磁共振确定。艾伦·布什博士是核磁共振分析多聚糖结构的主要专家。他的SubCore通过提供低聚糖、多糖、糖肽、小糖蛋白和糖脂的结构专业知识来支持聚乙二醇酯。最先进的核磁共振设备和方法在这个子核中可用。 次核C3-化学-Kevin Yarema博士是领先的化学糖生物学家。他的次级核心将通过以下方式支持挂钩：1)为“代谢性糖工程”提供糖类似物(例如。SCFA-己糖胺)； 2)执行抑制剂的定制合成(例如，D-PDMP、TMG)、叠氮糖和糖核苷酸、糖标记物和其他小分子。 次核心04-0-GlcNAc资源-Hart博士和Zachara博士是0-GlcNAc分析的领导者。 为了支持PEGS，我们提供：1)检测O-GlcNAc、OGT和OGA的工具；2)开发针对位点的O-GlcNAc抗体；3)为抗体生产和MS标准提供合成的O-GlcNAc多肽；4)OGT和OGA活性的分析；5)糖核苷酸和核苷酸的定量；6)协助单个蛋白质或全球蛋白质的位点定位和定量。这一共享资源提供了关键技术，以快速推进对NHLBI使命至关重要的糖科学研究。

PROJECT SU MMARY (See instructions): Analysis of glycoconjugate structure/functions requires specialized expertise from various disciplines. Many of these tools and approaches are not only difficult, but also require expensive and complex equipment. We have established our Shared Resource Core to not only contain the components that are most essential for the aims of this PEG, but also that build upon the unique strengths of our participating investigators. Each sub-Core is detailed in their separate sections. Sub-Core 01 - Mass Spectrometry -Drs. van Eyk and Zhang are leaders in mass spectrometric analysis of proteins and glycoproteins. The Core is extraordinarily well equipped. The Core's offerings include: 1) Identification of proteins; 2) global quantification of N-linked or O-linked glycoproteins using solidphase extraction of glycopeptides; 3) Lectin affinity/MS analyses; 4) Monitoring changes in glycosylation by lectin microarrays; 5) The use of "Click" chemistry for analyses of metabolically produced glycoconjugates; 6)Glycan profiling by MALDI-MS" (AXIMA-Resonance) and GlycanAnalyser. 7) Cell surface capture technology for isolation of glycoproteins; 2) Targeted capture of soluble glycoproteins from cytoplasm or extracellular environment; 3) MRM MS/MS assays for targeted quantification ofglycoproteins and glycosylation sites. Sub-Core C2 - NMR - When possible, glycan structure is best determined by NMR. Dr. Allen Bush is a leading expert in the structural analysis of glycans by NMR. His sub-Core supports the PEG by offering structural expertise for oligosaccharides, polysaccharides, glycopeptides, small glycoproteins and glycolipids. State of the art NMR equipment and methods are available in this sub-Core. Sub-Core C3 - Chemistry - Dr. Kevin Yarema is a leading chemical glycobiologist. His sub-Core will support the PEGs by: 1) providing sugar analogs for "metabolic glycoengineering" (eg. SCFA-hexosamines); 2) performing custom synthesis of inhibitors (eg. D-PDMP, TMG), azido sugars and sugar nucleotides, glycan-tagging tools and other small molecules. Sub-Core 04 - 0-GlcNAc Resources - Drs. Hart and Zachara are leaders in the analysis of 0-GlcNAc. In support of the PEGs we offer: 1) Tools for detecting 0-GlcNAc, OGT and OGA; 2) Develop site-specific 0-GlcNAc antibodies; 3) Provide synthetic 0-GlcNAc peptides for antibody production and MS standards; 4) Assays for OGT and OGA activity; 5) Quantitation of sugar nucleotides & nucleotides; 6) Assist with site mapping and quantification on single proteins or globally. This Shared Resource provides key technologies to allow rapid advancement of glycoscience research critical to the mission of the NHLBI.