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Title: Spin-Labeling Insights into How Chemical Fixation Impacts Glycan Organization on Cells
As new methods to interrogate glycan organization on cells develop, it is important to have a molecular level understanding of how chemical fixation can impact results and interpretations. Site-directed spin labeling technologies are well suited to study how the spin label mobility is impacted by local environmental conditions, such as those imposed by cross-linking effects of paraformaldehyde cell fixation methods. Here, we utilize three different azide-containing sugars for metabolic glycan engineering with HeLa cells to incorporate azido glycans that are modified with a DBCO-based nitroxide moiety via click reaction. Continuous wave X-band electron paramagnetic resonance spectroscopy is employed to characterize how the chronological sequence of chemical fixation and spin labeling impacts the local mobility and accessibility of the nitroxide-labeled glycans in the glycocalyx of HeLa cells. Results demonstrate that chemical fixation with paraformaldehyde can alter local glycan mobility and care should be taken in the analysis of data in any study where chemical fixation and cellular labeling occur.  more » « less
Award ID(s):
1715384
PAR ID:
10473165
Author(s) / Creator(s):
; ; ; ; ; ;
Corporate Creator(s):
Editor(s):
Zhongyu Yang
Publisher / Repository:
Springer
Date Published:
Journal Name:
Applied Magnetic Resonance
Edition / Version:
1
Volume:
x
Issue:
x
ISSN:
0937-9347
Page Range / eLocation ID:
x
Subject(s) / Keyword(s):
Sialic acid, metabolic glycan engineering, chemical fixation, crosslinking, glycan, organization, electron paramagnetic resonance spectroscopy
Format(s):
Medium: X Size: 1MB Other: pdf
Size(s):
1MB
Sponsoring Org:
National Science Foundation
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