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Identifying nonspecific ligand binding in electrospray ionization mass spectrometry using the reporter molecule method Open Access

Descriptions

Author or creator
Sun, Nian
Sun, Jiangxiao
Kitova, Elena N.
Klassen, John S.
Additional contributors
Subject/Keyword
Nucleic acids
Complexes
Constants
Ethidium-bromide
Duplex
Stability
Nanoelectrospray ionization
Dissociation
Affinities
DNA
Type of item
Journal Article (Published)
Language
English
Place
Time
Description
The application of the reporter molecule (Mrep) method for identifying nonspecific complexes in the ES-MS analysis of protein-ligand and DNA-ligand interactions in vitro is described. To test the reliability of the method, it was applied to the ES-MS analysis of protein-carbohydrate complexes originating from specific interactions in solution and from nonspecific interactions in the ES process. These control experiments confirm the basic assumptions underlying the Mrep method, namely that nonspecific ligand binding is a random process, and that the ES droplet histories for specific and nonspecific complexes are distinct. The application of the Mrep method to the ES-MS analysis of the sequential binding of the ethidium cation, a DNA intercalator, to single and double strand oligodeoxynucleotides is also described, and highlights the general utility of the method.
Date created
2009
DOI
doi:10.7939/R33B5WN5T
License information
© 2009 Sun, N., Sun, J., 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
Sun, N., Sun, J., Kitova, E. N., & Klassen, J. S. (2009). Identifying nonspecific ligand binding in electrospray ionization mass spectrometry using the reporter molecule method. Journal of the American Society for Mass Spectrometry, 20(7), 1242-1250.  http://doi.org/10.1016/j.jasms.2009.02.024

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