Comprehensive structural glycomic characterization of the glycocalyxes of cells and tissues

Nature Protocols
Qiongyu LiCarlito B Lebrilla

Abstract

The glycocalyx comprises glycosylated proteins and lipids and fcorms the outermost layer of cells. It is involved in fundamental inter- and intracellular processes, including non-self-cell and self-cell recognition, cell signaling, cellular structure maintenance, and immune protection. Characterization of the glycocalyx is thus essential to understanding cell physiology and elucidating its role in promoting health and disease. This protocol describes how to comprehensively characterize the glycocalyx N-glycans and O-glycans of glycoproteins, as well as intact glycolipids in parallel, using the same enriched membrane fraction. Profiling of the glycans and the glycolipids is performed using nanoflow liquid chromatography-mass spectrometry (nanoLC-MS). Sample preparation, quantitative LC-tandem MS (LC-MS/MS) analysis, and data processing methods are provided. In addition, we discuss glycoproteomic analysis that yields the site-specific glycosylation of membrane proteins. To reduce the amount of sample needed, N-glycan, O-glycan, and glycolipid analyses are performed on the same enriched fraction, whereas glycoproteomic analysis is performed on a separate enriched fraction. The sample preparation process takes 2-3 d, whereas the ti...Continue Reading

References

Jan 1, 1989·Advances in Cancer Research·S Hakomori
Sep 30, 1998·Proceedings of the National Academy of Sciences of the United States of America·H H ChouA Varki
Mar 30, 2002·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Sue Jackson, Susan W Nicolson
Aug 1, 1959·Canadian Journal of Biochemistry and Physiology·E G BLIGH, W J DYER
Sep 28, 2006·Journal of Agricultural and Food Chemistry·Milady R NinonuevoCarlito B Lebrilla
Jan 27, 2007·Pflügers Archiv : European journal of physiology·Sietze ReitsmaMirjam G A oude Egbrink
May 1, 2007·American Journal of Physiology. Heart and Circulatory Physiology·Yu YaoC Forbes Dewey
Sep 8, 2007·Molecular & Cellular Proteomics : MCP·Andreas J HülsmeierThierry Hennet
Dec 7, 2007·Mathematical Biosciences·Ryan S Senger, M Nazmul Karim
Mar 28, 2008·Analytical Chemistry·Richard R SeipertCarlito B Lebrilla
Sep 2, 2008·Journal of the American Society for Mass Spectrometry·Manolo D PlasenciaDavid E Clemmer
Jan 23, 2009·Journal of Hepatology·Bram BlommeHans Van Vlierberghe
Sep 25, 2009·Current Opinion in Chemical Biology·Hyun Joo AnCarlito B Lebrilla
Mar 13, 2010·Analytical and Bioanalytical Chemistry·L R RuhaakM Wuhrer
Jun 29, 2010·Journal of Proteome Research·Shuai WuCarlito B Lebrilla
Feb 11, 2012·Journal of Proteome Research·Danielle AldredgeCarlito B Lebrilla
Feb 23, 2012·Ageing Research Reviews·Fabio Dall'OlioClaudio Franceschi
Jun 26, 2012·The EMBO Journal·Peer-Hendrik KuhnStefan F Lichtenthaler
Oct 6, 2012·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yi DuHongcheng Liu
Oct 29, 2013·Analytical Biochemistry·Shisheng SunHui Zhang
Mar 22, 2014·Chemical Communications : Chem Comm·Van N Tra, Danielle H Dube
Jul 18, 2014·Nature·Matthew J PaszekValerie M Weaver
Dec 31, 2014·Proceedings of the National Academy of Sciences of the United States of America·Annie N SamrajAjit Varki
Jan 13, 2015·Journal of Autoimmunity·Emanual MaverakisCarlito B Lebrilla
Sep 1, 2015·Journal of Proteome Research·L Renee RuhaakSuzanne Miyamoto
May 4, 2016·Expert Review of Proteomics·Li CaoSi Wu
Jul 9, 2016·Scientific Reports·Narine ArabyanBart C Weimer

❮ Previous
Next ❯

Citations

Sep 18, 2020·Molecular & Cellular Proteomics : MCP·Nicholas M RileySharon J Pitteri
Feb 16, 2021·Molecular & Cellular Proteomics : MCP·Nicholas M RileySharon J Pitteri
Apr 4, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Byeong Gwan ChoYehia Mechref
Apr 9, 2021·Pflügers Archiv : European journal of physiology·Maia I KellyRebekah L Gundry
Mar 27, 2021·Metallomics : Integrated Biometal Science·Liming WangChunying Chen
Aug 3, 2021·Frontiers in Cell and Developmental Biology·Nuria VillalbaSarah Y Yuan
Aug 7, 2021·Molecular & Cellular Proteomics : MCP·Mariana BarbozaCarlito B Lebrilla
Sep 9, 2021·Mass Spectrometry Reviews·Guinevere S M Lageveen-KammeijerManfred Wuhrer

❮ Previous
Next ❯

Methods Mentioned

BETA
electrophoresis
chips
chip
glycosylation
peptide assay
Protein Assay
PCR
dissection
the

Software Mentioned

Byonic Viewer
Agilent MassHunter Qualitative Analysis
MassHunter
pGlyco
Windows
MassHunter Qualitative Analysis
Excel
ChipCube
Skyline
MaxQuant

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

© 2021 Meta ULC. All rights reserved