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Permanent link (DOI): https://doi.org/10.7939/R3WS8J08C

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Quantifying protein-carbohydrate interactions using liquid sample desorption electrospray ionization mass spectrometry Open Access

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Author or creator
Yao, Yuyu
Shams-Ud-Doha, Km
Daneshfar, Rambod
N. Kitova, Elena N.
Klassen, John S.
Additional contributors
Subject/Keyword
Liquid sample desorption electrospray ionization mass spectrometry
Affinity
Carbohydrate
Protein
Type of item
Journal Article (Published)
Language
English
Place
Time
Description
The application of liquid sample desorption electrospray ionization mass spectrometry (liquid sample DESI-MS) for quantifying protein–carbohydrate interactions in vitro is described. Association constants for the interactions between lysozyme and β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-D-GlcNAc and β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-D-GlcNAc, and between a single chain antibody and α-D-Galp-(1 → 2)-[α-D-Abep-(1 → 3)]-α-D-Manp-OCH3 and β-D-Glcp-(1 → 2)-[α-D-Abep-(1 → 3)]-α-D-Manp-OCH3 measured using liquid sample DESI-MS were found to be in good agreement with values measured by isothermal titration calorimetry and the direct ESI-MS assay. The reference protein method, which was originally developed to correct ESI mass spectra for the occurrence of nonspecific ligand-protein binding, was shown to reliably correct liquid sample DESI mass spectra for nonspecific binding. The suitability of liquid sample DESI-MS for quantitative binding measurements carried out using solutions containing high concentrations of the nonvolatile biological buffer phosphate buffered saline (PBS) was also explored. Binding of lysozyme to β-D-GlcNAc-(1 → 4)-β-D-GlcNAc-(1 → 4)-D-GlcNAc in aqueous solutions containing up to 1× PBS was successfully monitored using liquid sample DESI-MS; with ESI-MS the binding measurements were limited to concentrations less than 0.02 X PBS.
Date created
2015
DOI
doi:10.7939/R3WS8J08C
License information
© 2015 Yao, Y., Shams-Ud-Doha, K., Daneshfar, R., Kitova, E. N., & Klassen, J. S. This version of this article is open access and can be downloaded and shared. The original author(s) and source must be cited.
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Citation for previous publication
Yao, Y., Shams-Ud-Doha, K., Daneshfar, R., Kitova, E. N., & Klassen, J. S. (2015). Quantifying protein-carbohydrate interactions using liquid sample desorption electrospray ionization mass spectrometry. Journal of the American Society for Mass Spectrometry, 26(1), 98-106.  http://doi.org/10.1007/s13361-014-1008-7

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