Identification of potential sialic acid binding proteins on cell membranes by proximity chemical labeling

Chemical Science
Qiongyu LiCarlito B Lebrilla

Abstract

The cell membrane contains a highly interactive glycan surface on a scaffold of proteins and lipids. Sialic acids are negatively charged monosaccharides, and the proteins that bind to sialic acids play an important role in maintaining the integrity and collective functions of this interactive space. Sialic acid binding proteins are not readily identified and have nearly all been discovered empirically. In this research, we developed a proximity labeling method to characterize proteins with oxidation by localized radicals produced in situ. The sites of oxidation were identified and quantified using a standard proteomic workflow. In this method, a clickable probe was synthesized and attached to modified sialic acids on the cell membrane, which functioned as a catalyst for the localized formation of radicals from hydrogen peroxide. The proteins in the sialic acid environment were labeled through amino acid oxidation, and were categorized into three groups including sialylated proteins, non-sialylated proteins with transmembrane domains, and proteins that are associated with the membrane with neither sialylated nor transmembrane domains. The analysis of the last group of proteins showed that they were associated with binding functi...Continue Reading

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Citations

Aug 16, 2019·Cells·Qiongyu LiCarlito B Lebrilla
Nov 5, 2020·Mass Spectrometry Reviews·Xiao Pan, Richard W Vachet
Dec 5, 2020·Science·Dionna M KasperStefania Nicoli
May 29, 2021·Mass Spectrometry Reviews·Fangxu SunRonghu Wu
Aug 1, 2021·Proceedings of the National Academy of Sciences of the United States of America·Andrew L WolfeSung Eun Kim
Oct 19, 2019·Analytical Chemistry·Suttipong SuttapitugsakulRonghu Wu
Oct 7, 2021·Electrophoresis·Cristian D Gutierrez-ReyesYehia Mechref

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Methods Mentioned

BETA
footprinting
glycosylation
column chromatography
acetylation
confocal microscopy
Fluorescence imaging

Software Mentioned

POSE
Visual Molecular Dynamics ( VMD )
Byonic
MassHunter
MzIdentML
TMHMM
PANTHER
Skyline

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