Enzymatic Tagging of Glycoproteins on the Cell Surface for Their Global and Site-Specific Analysis with Mass Spectrometry

Analytical Chemistry
Fangxu SunRonghu Wu

Abstract

The cell surface is normally covered with sugars that are bound to lipids or proteins. Surface glycoproteins play critically important roles in many cellular events, including cell-cell communications, cell-matrix interactions, and response to environmental cues. Aberrant protein glycosylation on the cell surface is often a hallmark of human diseases such as cancer and infectious diseases. Global analysis of surface glycoproteins will result in a better understanding of glycoprotein functions and the molecular mechanisms of diseases and the discovery of surface glycoproteins as biomarkers and drug targets. Here, an enzyme is exploited to tag surface glycoproteins, generating a chemical handle for their selective enrichment prior to mass spectrometric (MS) analysis. The enzymatic reaction is very efficient, and the reaction conditions are mild, which are well-suited for surface glycoprotein tagging. For biologically triplicate experiments, on average 953 N-glycosylation sites on 393 surface glycoproteins per experiment were identified in MCF7 cells. Integrating chemical and enzymatic reactions with MS-based proteomics, the current method is highly effective to globally and site-specifically analyze glycoproteins only located on ...Continue Reading

References

May 1, 1990·The Journal of Membrane Biology·G von Heijne
Apr 8, 2000·Current Opinion in Structural Biology·M C Deller, E Yvonne Jones
Sep 21, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·M MartinP Engel
Nov 13, 2002·Plant Physiology·Andreas NebenführDavid G Robinson
Jan 5, 2005·Nature Reviews. Cancer·Fu-Tong Liu, Gabriel A Rabinovich
Jul 11, 2006·The Journal of Biological Chemistry·Laurence DuchesneDavid G Fernig
Feb 14, 2007·Proceedings of the National Academy of Sciences of the United States of America·Tsui-Ling HsuChi-Huey Wong
May 8, 2007·Nucleic Acids Research·Lukas KällErik L L Sonnhammer
Oct 19, 2007·Proceedings of the National Academy of Sciences of the United States of America·Jeremy M BaskinCarolyn R Bertozzi
Jan 10, 2009·Nature Protocols·Da Wei HuangRichard A Lempicki
Apr 15, 2009·Nature Reviews. Immunology·Gabriel A Rabinovich, Marta A Toscano
Feb 25, 2010·Nature Reviews. Cancer·Elena B Pasquale
Apr 13, 2010·Molecular & Cellular Proteomics : MCP·Rikke Leth-LarsenHenrik J Ditzel
Dec 25, 2010·Cell·Edward L HuttlinSteven P Gygi
May 28, 2011·Current Opinion in Cell Biology·Cecile BoscherIvan R Nabi
Jul 30, 2011·Bioorganic & Medicinal Chemistry Letters·Sarah C HubbardCarolyn R Bertozzi
Oct 1, 2011·Nature Methods·Thomas Nordahl PetersenHenrik Nielsen
Mar 23, 2012·Nature Reviews. Cancer·Andrew M ScottLloyd J Old
Oct 17, 2012·Glycobiology·T N C RamyaJames C Paulson
May 24, 2013·Nature Reviews. Molecular Cell Biology·Federica Brandizzi, Charles Barlowe
Jul 23, 2013·Nature Chemical Biology·Markus Grammel, Howard C Hang
Sep 14, 2013·Science·Mark X Sliwkowski, Ira Mellman
Nov 1, 1994·Journal of the American Society for Mass Spectrometry·J K EngJ R Yates
Apr 3, 2014·Molecular & Cellular Proteomics : MCP·Weixuan ChenRonghu Wu
Sep 23, 2014·Nature Reviews. Immunology·Matthew S MacauleyJames C Paulson
Nov 27, 2014·Journal of the American Society for Mass Spectrometry·Johanna M SmeekensRonghu Wu
Jan 27, 2015·Annual Review of Pathology·Sean R StowellRichard D Cummings
Apr 22, 2015·PloS One·Damaris Bausch-FluckBernd Wollscheid
Jun 14, 2015·Annual Review of Analytical Chemistry·Zhikai Zhu, Heather Desaire
Feb 20, 2016·Analytical Chemistry·Haopeng XiaoRonghu Wu
Jun 16, 2016·Proteomics·Tujin ShiRichard D Smith
Aug 18, 2016·Journal of Proteome Research·Lauren G ZachariasYehia Mechref
Oct 22, 2016·Cell·Erik McShaneMatthias Selbach
May 18, 2017·Angewandte Chemie·Jiangnan ZhengRonghu Wu

❮ Previous
Next ❯

Citations

Sep 15, 2019·Journal of Separation Science·Yanan LiMingliang Ye
Jul 30, 2019·Expert Review of Proteomics·Amanda HaymondVirginia Espina
Sep 18, 2020·Molecular & Cellular Proteomics : MCP·Nicholas M RileySharon J Pitteri
Mar 5, 2021·Analytical Chemistry·Yanan LiMingliang Ye
Feb 16, 2021·Molecular & Cellular Proteomics : MCP·Nicholas M RileySharon J Pitteri
Apr 17, 2021·Biochemical Society Transactions·Kathryn E Huxley, Lianne I Willems
May 29, 2021·Mass Spectrometry Reviews·Fangxu SunRonghu Wu
Oct 19, 2019·Analytical Chemistry·Suttipong SuttapitugsakulRonghu Wu

❮ Previous
Next ❯

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.