Matrix assisted laser desorption ionization imaging mass spectrometry workflow for spatial profiling analysis of N-linked glycan expression in tissues.
Matrix assisted laser desorption ionization imaging mass spectrometry workflow for spatial profiling analysis of N-linked glycan expression in tissues.
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DOI：
10.1021/ac402108x
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发表时间：
2013-10-15
影响因子：
7.4
通讯作者：
Drake, Richard R.
中科院分区：
文献类型：
--
作者：
Powers, Thomas W.;Jones, E. Ellen;Betesh, Lucy R.;Romano, Patrick R.;Gao, Peng;Copland, John A.;Mehta, Anand S.;Drake, Richard R.
描述了一种新的基质辅助激光解吸电离成像质谱学(MALDI-IMS)方法，用于组织中多个N-连接的糖链物种的位置和分布。直接在组织上应用一种内切糖苷酶，即多肽N-糖苷酶F(PNGaseF)，然后孵育释放出适合MALDI-IMS检测的N-连接的糖链种类。该方法被设计为同时分析从细胞内细胞器和细胞表面糖蛋白释放的多种糖类，同时保持组织病理学兼容的制备工作流程。将重组PNGaseF酶均匀喷洒在小鼠脑组织切片上，孵育2小时，然后喷洒2，5-二羟基苯甲酸基质进行MALDI-IMS分析。使用这一基本方法，检测了主要细胞N-连接糖体的全球快照，包括它们的组织定位和分布、结构和相对丰度。从相似加工的组织中进行组织外提取和修饰，并进行进一步的质谱分析或高效液相色谱分析以指定结构名称。MALDI-IMS主要用于组织中蛋白质、脂类、药物和小分子代谢物的空间图谱分析，但尚未被应用于N-连接的糖链分析。本文所述的可翻译的MALDI-IMS葡聚糖谱工作流程可以容易地应用于任何感兴趣的组织类型。从临床诊断学的角度来看，区分N-糖链并将其分子表达与组织病理学变化相关联的能力可以为识别生物标记物和治疗应用的新的疾病相关靶点提供新的方法。
A new Matrix Assisted Laser Desorption Ionization Imaging Mass Spectrometry (MALDI-IMS) method to spatially profile the location and distribution of multiple N-linked glycan species in tissues is described. Application of an endoglycosidase, peptide N-glycosidase F (PNGaseF), directly on tissues followed by incubation releases N-linked glycan species amenable to detection by MALDI-IMS. The method has been designed to simultaneously profile the multiple glycan species released from intracellular organelle and cell surface glycoproteins, while maintaining histopathology compatible preparation workflows. A recombinant PNGaseF enzyme was sprayed uniformly across mouse brain tissue slides, incubated for two hours, then sprayed with 2,5-dihydroxybenzoic acid matrix for MALDI-IMS analysis. Using this basic approach, global snapshots of major cellular N-linked glycoforms were detected, including their tissue localization and distribution, structure and relative abundance. Off-tissue extraction and modification of glycans from similarly processed tissues and further mass spectrometry or HPLC analysis was done to assign structural designations. MALDI-IMS has primarily been utilized to spatially profile proteins, lipids, drug and small molecule metabolites in tissues, but it has not been previously applied to N-linked glycan analysis. The translatable MALDI-IMS glycan profiling workflow described herein can readily be applied to any tissue type of interest. From a clinical diagnostics perspective, the ability to differentially profile N glycans and correlate their molecular expression to histopathological changes can offer new approaches to identifying novel disease related targets for biomarker and therapeutic applications.
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通过质谱法对组织中脂质生物化学的MALDI成像。
影响因子：
62.1
作者：
Berry, Karin A. Zemski;Hankin, Joseph A.;Barkley, Robert M.;Spraggins, Jeffrey M.;Caprioli, Richard M.;Murphy, Robert C.
通讯作者：
Murphy, Robert C.
影响因子：
64.5
作者：
Dennis JW;Nabi IR;Demetriou M
通讯作者：
Demetriou M
DOI：
10.1155/2012/560391
发表时间：
2012
期刊：
International journal of proteomics
影响因子：
--
作者：
Saba J;Dutta S;Hemenway E;Viner R
通讯作者：
Viner R
影响因子：
7
作者：
Chaurand, Pierre;Cornett, Dale S.;Caprioli, Richard M.
通讯作者：
Caprioli, Richard M.
影响因子：
3.7
作者：
Nilsson A;Fehniger TE;Gustavsson L;Andersson M;Kenne K;Marko-Varga G;Andrén PE
通讯作者：
Andrén PE