Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis

Mélissa Robb; Joanne K Hobbs; Alisdair B Boraston

doi:10.1007/978-1-4939-6899-2_17

Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis

Mélissa Robb, Joanne K Hobbs, Alisdair B Boraston

School of Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Fluorophore-assisted carbohydrate electrophoresis (FACE) is a method in which a fluorophore is covalently attached to the reducing end of carbohydrates, thereby allowing visualization following high-resolution separation by electrophoresis. This method can be used for carbohydrate profiling and sequencing, as well as for the determination of the specificity of carbohydrate-active enzymes. Here, we describe and demonstrate the use of FACE to separate and visualize the glycans released following digestion of oligosaccharides by glycoside hydrolases (GHs) using two examples: (1) the digestion of chitobiose by the streptococcal β-hexosaminidase GH20C, and (2) the digestion of glycogen by the GH13 member SpuA.

Original language	English
Pages (from-to)	215-221
Number of pages	7
Journal	Methods in Molecular Biology
Volume	1588
DOIs	https://doi.org/10.1007/978-1-4939-6899-2_17
Publication status	Published - 2017

Keywords

Carbohydrates/analysis
Disaccharides/analysis
Electrophoresis/methods
Glycogen/analysis
Glycoside Hydrolases/metabolism
Pyrenes/chemistry

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10.1007/978-1-4939-6899-2_17

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title = "Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis",

abstract = "Fluorophore-assisted carbohydrate electrophoresis (FACE) is a method in which a fluorophore is covalently attached to the reducing end of carbohydrates, thereby allowing visualization following high-resolution separation by electrophoresis. This method can be used for carbohydrate profiling and sequencing, as well as for the determination of the specificity of carbohydrate-active enzymes. Here, we describe and demonstrate the use of FACE to separate and visualize the glycans released following digestion of oligosaccharides by glycoside hydrolases (GHs) using two examples: (1) the digestion of chitobiose by the streptococcal β-hexosaminidase GH20C, and (2) the digestion of glycogen by the GH13 member SpuA.",

keywords = "Carbohydrates/analysis, Disaccharides/analysis, Electrophoresis/methods, Glycogen/analysis, Glycoside Hydrolases/metabolism, Pyrenes/chemistry",

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year = "2017",

doi = "10.1007/978-1-4939-6899-2_17",

language = "English",

volume = "1588",

pages = "215--221",

journal = "Methods in Molecular Biology",

issn = "1064-3745",

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T1 - Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis

AU - Robb, Mélissa

AU - Hobbs, Joanne K

AU - Boraston, Alisdair B

PY - 2017

Y1 - 2017

N2 - Fluorophore-assisted carbohydrate electrophoresis (FACE) is a method in which a fluorophore is covalently attached to the reducing end of carbohydrates, thereby allowing visualization following high-resolution separation by electrophoresis. This method can be used for carbohydrate profiling and sequencing, as well as for the determination of the specificity of carbohydrate-active enzymes. Here, we describe and demonstrate the use of FACE to separate and visualize the glycans released following digestion of oligosaccharides by glycoside hydrolases (GHs) using two examples: (1) the digestion of chitobiose by the streptococcal β-hexosaminidase GH20C, and (2) the digestion of glycogen by the GH13 member SpuA.

AB - Fluorophore-assisted carbohydrate electrophoresis (FACE) is a method in which a fluorophore is covalently attached to the reducing end of carbohydrates, thereby allowing visualization following high-resolution separation by electrophoresis. This method can be used for carbohydrate profiling and sequencing, as well as for the determination of the specificity of carbohydrate-active enzymes. Here, we describe and demonstrate the use of FACE to separate and visualize the glycans released following digestion of oligosaccharides by glycoside hydrolases (GHs) using two examples: (1) the digestion of chitobiose by the streptococcal β-hexosaminidase GH20C, and (2) the digestion of glycogen by the GH13 member SpuA.

KW - Carbohydrates/analysis

KW - Disaccharides/analysis

KW - Electrophoresis/methods

KW - Glycogen/analysis

KW - Glycoside Hydrolases/metabolism

KW - Pyrenes/chemistry

U2 - 10.1007/978-1-4939-6899-2_17

DO - 10.1007/978-1-4939-6899-2_17

M3 - Article

C2 - 28417372

SN - 1064-3745

VL - 1588

SP - 215

EP - 221

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -

Separation and Visualization of Glycans by Fluorophore-Assisted Carbohydrate Electrophoresis

Abstract

Keywords

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