Advancing Ultraviolet Photodissociation Mass Spectrometry for Precision Mapping of Protein Glycosylation
Advancing Ultraviolet Photodissociation Mass Spectrometry for Precision Mapping of Protein Glycosylation
批准号:
10350600
负责人:
Edwin E Escobar
金额:
$2.77万
依托单位国家:
美国
项目类别:
财政年份:
2021
资助国家:
美国
项目状态:
已结题
起止时间:
2021-01-01 至 2022-07-31
关键词:
Amino AcidsAtherosclerosisBiochemicalBiologicalC-terminalCell CommunicationCell membraneCell physiologyCellsChemicalsCodeComplexConsensusConsensus SequenceCoupledDefectDevelopmentDiseaseDisease PathwayDissociationEnzymesEpigenetic ProcessEukaryotaEventFunctional disorderGene ProteinsGlycopeptidesGlycoproteinsGlycosidesGoalsGoldHealthHumanImmune responseImmunityIonsKnowledgeLengthLightLinkLocationMalignant NeoplasmsMapsMass Spectrum AnalysisMediatingMessenger RNAMetabolismMethodologyMethodsModificationN-Glycosylation SiteNatureNeoplasm MetastasisPathogenicityPathway interactionsPatternPeptidesPhosphorylationPhotonsPlayPolysaccharidesPositioning AttributePost Translational Modification AnalysisPost-Translational Protein ProcessingProcessProlineProtein GlycosylationProtein-Serine-Threonine KinasesProteinsProteomeProteomicsRNA Polymerase IIRegulationReportingResearchResolutionRoleSamplingSerineSignal TransductionSiteStructureStructure-Activity RelationshipTechniquesThreonineThrombosisTyrosineViralViral Envelope ProteinsVirus Diseasesanalytical methodanalytical toolbaseglycoproteomicsglycosylationhuman diseaseinnovationinsightpolysulfated glycosaminoglycansugartandem mass spectrometrytooltumorultraviolet
中文摘要
项目摘要
糖基化在细胞中蛋白质的所有基本特征中起着结构和功能作用。尽管作出了重大的
有证据表明O-糖基化在细胞过程中至关重要,O-连接聚糖的性质和特定位置
O-聚糖的特性还没有得到很好的表征。在很大程度上,共有序列基序N-X-S/T使得能够可靠地识别出与N-X-S/T相关的基因。
预测的N-糖基化位点,但我们的知识的O-糖基化是阻碍了缺乏简单的共识基序。
因此,我们目前对O-糖基化的影响的理解是不完整的。全面执行《公约》的主要障碍
通过基于质谱的蛋白质组学工作流程表征O-糖基化是
不稳定的O-连接的聚糖使用传统的碰撞活化解离,妨碍精确定位。更没有
存在可切割每种可能的O-聚糖β-O-糖苷键的全局酶,从而阻碍酶促糖组学分析。
与紫外光解离(UVPD)耦合的质谱被定位为是质谱分析的重要工具。
通过实现与人类健康有关的蛋白质的PTM的残基水平分辨率,可以实现糖蛋白质组学。UVPD可以是
同时提供肽的高序列覆盖率和不稳定修饰的保留,
允许O-聚糖作图和结构表征。我开发的创新LC-UVPD-MS策略适用于
对于靶向和全局糖蛋白质组学应用,
疾病的途径,并推动新的生物学问题。
英文摘要
PROJECT SUMMARY
Glycosylation plays a structural and functional role in all fundamental features of proteins in cells. Despite substantial
evidence suggesting O-glycosylation is vital in cellular processes, the nature of O-linked glycans and the specific locations
of O-glycans are not well characterized. To a large extent, the consensus sequence motif N-X-S/T enables reliable
prediction of N-glycosylation sites, but our knowledge of O-glycosylation is hampered by a lack of simple consensus motifs.
As a result, our current understanding of the impact of O-glycosylation is incomplete. A major obstacle to comprehensive
characterization of O-glycosylation by mass spectrometry-based proteomic workflows is the prominent neutral loss of
labile O-linked glycans using conventional collisional activated dissociation, hampering precise localization. Moreover, no
global enzyme exists that can cleave every possible O-glycan β-O-glycosidic linkage, thwarting enzymatic glycomic analysis.
Mass spectrometry coupled with ultraviolet photodissociation (UVPD) is positioned to be an important tool in
glycoproteomics by enabling residue-level resolution of PTMs of proteins implicated in human health. UVPD can be
harnessed to provide simultaneously high sequence coverage of peptides and retention of labile modifications, thus
allowing O-glycan mapping and structural characterization. My development of innovative LC-UVPD-MS strategies suitable
for both targeted and global glycoproteomic applications will provide new insight into the correlation of glycosylation with
disease pathways and drive new biological questions.
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