ERA

Download the full-sized PDF of Screening oligosaccharide libraries against lectins using the proxy protein electrospray ionization mass spectrometry assayDownload the full-sized PDF

Analytics

Share

Permanent link (DOI): https://doi.org/10.7939/R3K931K9R

Download

Export to: EndNote  |  Zotero  |  Mendeley

Communities

This file is in the following communities:

Chemistry, Department of

Collections

This file is in the following collections:

Journal Articles (Chemistry)

Screening oligosaccharide libraries against lectins using the proxy protein electrospray ionization mass spectrometry assay Open Access

Descriptions

Author or creator
Li, Jun
Fan, Xuxin
Kitova, Elena N.
Zou, Chunxia
Cairo, Christopher W.
Eugenio, Luiz
Ng, Kenneth K. S.
Xiong, Zi Jian
Privé, Gilbert G.
Klassen, John S.
Additional contributors
Subject/Keyword
Drug discovery
Binding
Glycan microarrays
Carbohydrate ligands
Complexes
Frontal affinity-chromatography
Catch
Group antigen receptors
Blood-group antigens
P-domain
Type of item
Journal Article (Published)
Language
English
Place
Time
Description
An electrospray ionization mass spectrometry (ESI-MS) assay for screening carbohydrate libraries against lectins is described. The assay is based on the proxy protein ESI-MS method, which combines direct ESI-MS protein–ligand binding measurements and competitive protein binding, to simultaneously detect and quantify protein–carbohydrate interactions. Specific interactions between components of the library and the target protein (PT) are identified from changes in the relative abundances (as measured by ESI-MS) of the carbohydrate complexes of a proxy protein (Pproxy), which binds to all components of the library with known affinity, upon addition of PT to the solution. The magnitude of the change in relative abundance of a given Pproxy–ligand complex provides a quantitative measure of the affinity of the corresponding PT–ligand interaction. A mathematical framework for the implementation of the method in the case of monovalent (single binding site) Pproxy and monovalent and multivalent (multiple equivalent and independent binding sites) PT is described. The application of the method to screen small libraries of oligosaccharides, on the basis of human histo-blood group antigens and milk oligosaccharides, against an N-terminal fragment of the family 51 carbohydrate-binding module, a fucose-binding lectin from Ralstonia solanacearum, and human norovirus VA387 P particle (24-mer of the protruding domain of the capsid protein), serves to demonstrate the reliability and versatility of the assay.
Date created
2016
DOI
doi:10.7939/R3K931K9R
License information
© 2016 Han, L., Shams-Ud-Doha, K., 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.
Rights

Citation for previous publication
Han, L., Shams-Ud-Doha, K., Kitova, E. N., & Klassen, J. S. (2016). Screening oligosaccharide libraries against lectins using the proxy protein electrospray ionization mass spectrometry assay. Analytical Chemistry, 88(16), 8224-8231.  http://doi.org/10.1021/acs.analchem.6b02044

Source

Link to related item

File Details

Date Uploaded
Date Modified
Audit Status
Audits have not yet been run on this file.
Characterization
File format: pdf (Portable Document Format)
Mime type: application/pdf
File size: 2802232
Last modified: 2017:10:11 14:26:41-06:00
Filename: AC_88_16_8224.pdf
Original checksum: 79db83815fe5460bfb6d913da57645ab
Well formed: true
Valid: true
File author: Klassen Group
Page count: 51
File language: en-US
Activity of users you follow
User Activity Date