Quantifying carbohydrate-protein interactions by electrospray ionization mass spectrometry analysis

Biochemistry
Amr El-HawietJohn S Klassen

Abstract

The development of analytical methods capable of characterizing carbohydrate-protein interactions, which are critical for many biological processes, represents an active area of research. Recently, the direct electrospray ionization mass spectrometry (ESI-MS) assay has emerged as a valuable tool for identifying and quantifying carbohydrate-protein complexes in vitro. The assay boasts a number of strengths, including its simplicity, speed, low level of sample consumption, and the unique ability to directly probe binding stoichiometry and to measure multiple binding equilibria simultaneously. Here, we describe the implementation of the direct ESI-MS assay for the determination of carbohydrate-protein binding stoichiometries and affinities. Common sources of error encountered with direct ESI-MS analysis of carbohydrate-protein interactions are identified along with strategies for minimizing their effects. The application of ESI-MS and a catch-and-release strategy for carbohydrate library screening are also described. The utility of the direct ESI-MS assay can be extended by combining the technique with competitive protein or ligand binding. An overview of these "indirect" ESI-MS methods is given, as well as examples of recent appl...Continue Reading

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Citations

Apr 11, 2015·Journal of the American Chemical Society·Simon NgRatmir Derda
Jun 2, 2015·Methods : a Companion to Methods in Enzymology·Julien Marcoux, Sarah Cianférani
May 6, 2015·Glycobiology·Amr El-HawietJohn S Klassen
Oct 16, 2014·Journal of the American Society for Mass Spectrometry·Yuyu YaoJohn S Klassen
Dec 19, 2019·Molecular & Cellular Proteomics : MCP·Brian B Haab, Zachary Klamer
Aug 15, 2017·Rapid Communications in Mass Spectrometry : RCM·Marko Haramija, Jasna Peter-Katalinić
Jan 1, 2014·Bacteriophage·Denis ArutyunovChristine M Szymanski
Jan 17, 2021·Molecular & Cellular Proteomics : MCP·Brian B Haab, Zachary Klamer
Feb 26, 2021·Analytical Chemistry·Erick G Báez BolivarJohn S Klassen
Jun 9, 2017·Journal of the American Society for Mass Spectrometry·Katharina RootRenato Zenobi
Jan 10, 2020·ACS Chemical Biology·Alexander EddendenMark Nitz

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