LC/MS analysis of complex multiglycosylated human alpha(1)-acid glycoprotein as a model for developing identification and quantitation methods for intact glycopeptide analysis

Analytical Biochemistry
Melanie M IvancicMichael J Treuheit

Abstract

The site-specific characterization of the complex glycans in multiglycosylated proteins requires developing methods where the carbohydrates remain covalently bound to the protein. The complexity in the carbohydrate composition of alpha(1)-acid glycoprotein (AAG) makes it an ideal model protein for such development. AAG has five N-asparaginyl-linked glycosylation sites, each varying in its bi-, tri-, and tetraantennary glycan content. We present an on-line liquid chromatography/mass spectrometry (LC/MS) method that uses high-low cone voltage switching for in-source fragmentation to determine the structures of the complex glycans present on each site for the two gene products of AAG. High cone voltage caused carbohydrate fragmentation, leading to the generation of signature carbohydrate ions that we used as markers to identify the glycopeptides. Low cone voltage produced minimal carbohydrate fragmentation and enabled the identification and quantification of the intact oligosaccharide structures on each glycopeptide based on its monoisotopic mass and intensity. Quantitation was accomplished by using the intensities of peaks from deconvoluted and deisotoped mass spectra or from the areas of the extracted ion chromatograms from the ...Continue Reading

References

Sep 30, 1992·Clinica Chimica Acta; International Journal of Clinical Chemistry·M J Treuheit, H B Halsall
Apr 1, 1992·The Biochemical Journal·M J TreuheitH B Halsall
Sep 1, 1983·Canadian Journal of Biochemistry and Cell Biology = Revue Canadienne De Biochimie Et Biologie Cellulaire·H SchachterG Vella
Jan 1, 1982·Preparative Biochemistry·H B HalsallM L Friedman
Nov 4, 2000·Biochimica Et Biophysica Acta·T FournierD Porquet
Jun 21, 2003·Clinical Chemistry·Thomas M Annesley
Jan 18, 2006·Methods in Enzymology·Katalin F Medzihradszky
Apr 11, 2006·Journal of the American Society for Mass Spectrometry·Jennifer L SeymourJoseph Zaia
Nov 23, 2006·Biochimica Et Biophysica Acta·Bogdan A BudnikJudith A J Steen
Dec 1, 2006·Biochimica Et Biophysica Acta·Hildegard Geyer, Rudolf Geyer
Feb 20, 2007·Current Protein & Peptide Science·Fabrizio Ceciliani, Vanessa Pocacqua
Jul 17, 2007·Analytical Chemistry·Pilsoo KangMilos V Novotny
Aug 28, 2007·Journal of the American Society for Mass Spectrometry·Catherine E CostelloJohn F Cipollo

❮ Previous
Next ❯

Citations

Feb 23, 2012·Analytical and Bioanalytical Chemistry·John E SchielKaren W Phinney
Feb 9, 2013·BMC Medicine·Uros KuzmanovEleftherios P Diamandis
Mar 29, 2014·Journal of the American Society for Mass Spectrometry·Zhikai ZhuHeather Desaire
Apr 15, 2016·Scientific Reports·Bas C JansenManfred Wuhrer
Jul 15, 2011·Journal of Pharmaceutical and Biomedical Analysis·Song C KlapoetkeSteven Becht
Apr 2, 2013·Advanced Drug Delivery Reviews·Eric Ezan
Oct 10, 2013·Acta Biochimica Et Biophysica Sinica·Shu ZhangYinkun Liu
Dec 1, 2015·Scientific Reports·Shuang YangShuwei Li
Feb 23, 2012·Annals of the New York Academy of Sciences·Eugene Dempsey, Pauline M Rudd
Apr 30, 2014·The Analyst·S DefausR Gutiérrez-Gallego

❮ 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.