Glycomic Profiling of Cells and Tissues by Mass Spectrometry: Fingerprinting and Sequencing Methodologies

Over the past decade, rapid, high‐sensitivity mass spectrometric strat‐egies have been developed and optimized for screening for the types of N‐ and O‐glycans present in a diverse range of biological material, including secretions, cell lines, tissues, and organs. These glycomic strategies are based...

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Veröffentlicht in:Methods in Enzymology 2006, Vol.415, p.59-86
Hauptverfasser: Jang‐Lee, Jihye, North, Simon J., Sutton‐Smith, Mark, Goldberg, David, Panico, Maria, Morris, Howard, Haslam, Stuart, Dell, Anne
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container_start_page 59
container_title Methods in Enzymology
container_volume 415
creator Jang‐Lee, Jihye
North, Simon J.
Sutton‐Smith, Mark
Goldberg, David
Panico, Maria
Morris, Howard
Haslam, Stuart
Dell, Anne
description Over the past decade, rapid, high‐sensitivity mass spectrometric strat‐egies have been developed and optimized for screening for the types of N‐ and O‐glycans present in a diverse range of biological material, including secretions, cell lines, tissues, and organs. These glycomic strategies are based on matrix‐assisted laser desorption/ionization (MALDI) time‐of‐flight mass fingerprinting of permethylated derivatives, combined with electrospray (ES) or MALDI tandem mass spectrometry (MS/MS) sequencing and gas chromatography (GC)‐MS linkage analysis, complemented by chemical and enzymatic degradations. Protocols for these methods are described in the first part of this chapter. Glycomic experiments yield large volumes of MS data, and interpretation of the resulting spectra remains a time‐consuming bottleneck in the process. In the second part of this chapter, we describe the use and operation of a mass spectral viewer program capable of displaying and automatically labeling spectra arising from MALDI fingerprinting of N‐glycans.
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subjects Amino Acid Sequence
Animals
Carbohydrate Conformation
Carbohydrate Sequence
Glycoproteins - chemistry
Humans
Mice
Molecular Sequence Data
Peptides - chemistry
Peptides - metabolism
Polysaccharides - chemistry
Proteomics - methods
Software
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods
title Glycomic Profiling of Cells and Tissues by Mass Spectrometry: Fingerprinting and Sequencing Methodologies
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