Quantifying Protein-Carbohydrate Interactions Using Liquid Sample Desorption Electrospray Ionization Mass Spectrometry

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-...

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Bibliographic Details
Published in:Journal of the American Society for Mass Spectrometry 2015-01, Vol.26 (1), p.98-106
Main Authors: Yao, Yuyu, Shams-Ud-Doha, Km, Daneshfar, Rambod, Kitova, Elena N., Klassen, John S.
Format: Article
Language:English
Subjects:
Analytical Chemistry
Binding
Bioinformatics
Biotechnology
Buffers
Carbohydrates
Carbohydrates - chemistry
Chemistry
Chemistry and Materials Science
Desorption
Electrospraying
Heat measurement
Ionization
Ions
Lysozyme
Mass spectra
Mass spectrometry
Mass spectroscopy
Models, Chemical
Organic Chemistry
Protein Binding
Proteins
Proteins - chemistry
Proteins - metabolism
Proteomics
Research Article
Scientific imaging
Sodium Chloride
Spectrometry, Mass, Electrospray Ionization - methods
Titration calorimetry
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Summary: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-OCH 3 and β-D-Glcp-(1 → 2)-[α-D-Abep-(1 → 3)]-α-D-Manp-OCH 3 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. Graphical Abstract ᅟ
ISSN:1044-0305
1879-1123
DOI:10.1007/s13361-014-1008-7