ERA

Download the full-sized PDF of Detecting protein-glycolipid interactions using glycomicelles and CaR-ESI-MSDownload the full-sized PDF

Analytics

Share

Permanent link (DOI): https://doi.org/10.7939/R3639KK0T

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)

Detecting protein-glycolipid interactions using glycomicelles and CaR-ESI-MS Open Access

Descriptions

Author or creator
Han, Ling
Kitova, Elena N.
Klassen, John S.
Additional contributors
Subject/Keyword
Mass spectrometry
Electrospray ionization
Interactions
Protein
Glycolipid
Micelles
Affinity
Type of item
Journal Article (Published)
Language
English
Place
Time
Description
This study reports on the use of the catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay, combined with glycomicelles, as a method for detecting specific interactions between water-soluble proteins and glycolipids (GLs) in aqueous solution. The B subunit homopentamers of cholera toxin (CTB5) and Shiga toxin type 1 B (Stx1B5) and the gangliosides GM1, GM2, GM3, GD1a, GD1b, GT1b, and GD2 served as model systems for this study. The CTB5 exhibits broad specificity for gangliosides and binds to GM1, GM2, GM3, GD1a, GD1b, and GT1b; Stx1B5 does not recognize gangliosides. The CaR-ESI-MS assay was used to analyze solutions of CTB5 or Stx1B5 and individual gangliosides (GM1, GM2, GM3, GD1a, GD1b, GT1b, and GD2) or mixtures thereof. The high affinity interaction of CTB5 with GM1 was successfully detected. However, the apparent affinity, as determined from the mass spectra, is significantly lower than that of the corresponding pentasaccharide or when GM1 is presented in model membranes such as nanodiscs. Interactions between CTB5 and the low affinity gangliosides GD1a, GD1b, and GT1b, as well as GD2, which served as a negative control, were detected; no binding of CTB5 to GM2 or GM3 was observed. The CaR-ESI-MS results obtained for Stx1B5 reveal that nonspecific protein-ganglioside binding can occur during the ESI process, although the extent of binding varies between gangliosides. Consequently, interactions detected for CTB5 with GD1a, GD1b, and GT1b are likely nonspecific in origin. Taken together, these results reveal that the CaR-ESI-MS/glycomicelle approach for detecting protein–GL interactions is prone to false positives and false negatives and must be used with caution.
Date created
2016
DOI
doi:10.7939/R3639KK0T
License information
© 2016 Han, L., 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., Kitova, E. N., & Klassen, J. S. (2016). Detecting protein-glycolipid interactions using glycomicelles and CaR-ESI-MS. Journal of the American Society for Mass Spectrometry, 27(11), 1878-1886.  http://dx.doi.org/10.1007/s13361-016-1461-6

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: 4686757
Last modified: 2017:10:11 14:27:06-06:00
Filename: JASMS_27_11_1878.pdf
Original checksum: 1d82bc99436ae0bb83589cc37e8e4e92
Well formed: true
Valid: true
File author: Sherry
Page count: 59
File language: en-US
Activity of users you follow
User Activity Date