Skip to main content
Skip to main menu Skip to spotlight region Skip to secondary region Skip to UGA region Skip to Tertiary region Skip to Quaternary region Skip to unit footer

Slideshow

Negative Electron Transfer Dissociation paired with Capillary Electrophoresis-Mass Spectrometry for the investigation of Glycosaminoglycan mixtures

Morgan Stickney
Morgan Stickney
Chemistry Department
University of Georgia
Chemistry Building, Room 400
Analytical Seminar

Sulfated glycosaminoglycan (GAG) carbohydrates are long, linear, acidic polysaccharide chains abundant on the surface of virtually all mammalian cells[1,2]. Non-template driven modifications affect many biological functions through protein-binding interactions[3]. The complex structure and low natural abundance of GAG oligomers remains a significant analytical challenge. Capillary electrophoresis-mass spectrometry (CE-MS) presents an efficient approach for online separation of complex GAG mixtures when paired with a commercial interface that has a low dilution factor. Mixtures of purified GAG standards have been baseline resolved and biological mixtures have been significantly reduced in complexity[4].

Tandem mass spectrometry is essential for characterization of the modification patterns and binding motifs of GAGs. Negative Electron Transfer Dissociation (NETD) is a powerful fragmentation method that can be performed on mass spectrometry platforms capable of ion-ion reactions. Electron based fragmentation methods like NETD cause radical rearrangement reactions that fragment GAG backbones, without stripping sensitive sulfo-modifications, on a time scale amenable to CE separations[5]. By combining a CE-MS separation platform with NETD we can obtain precise structural characterization of diasteriomeric GAG mixtures to determine binding motifs responsible for GAG chain interactions with proteins. Future research will see this platform applied to protein pulldown samples, and automation of sample analysis.

(1) Kailemia, M. J., et al. (2014). "Oligosaccharide Analysis By Mass Spectrometry: A Review Of Recent Developments." Analytical Chemistry 86(1): 196-212.

(2) Laremore, T. N., et al. (2010). "High-resolution preparative separation of glycosaminoglycan oligosaccharides by polyacrylamide gel electrophoresis." Analytical Biochemistry 401(2): 236-241.

(3)Aquino, R. S. and P. W. Park (2016). "Glycosaminoglycans and infection." Frontiers in bioscience (Landmark edition) 21: 1260-1277.

(4) Sanderson, P., et al. (2018). "Heparin/heparan sulfate analysis by covalently modified reverse polarity capillary zone electrophoresis-mass spectrometry." Journal of Chromatography A 1545: 75-83.

(5) Leach, F. E., et al. (2017). "Negative Electron Transfer Dissociation Sequencing of Increasingly Sulfated Glycosaminoglycan Oligosaccharides on an Orbitrap Mass Spectrometer." Journal of The American Society for Mass Spectrometry 28(9): 1844-1854.

Support Us

We appreciate your financial support. Your gift is important to us and helps support critical opportunities for students and faculty alike, including lectures, travel support, and any number of educational events that augment the classroom experience. Click here to learn more about giving.

Every dollar given has a direct impact upon our students and faculty.

Got More Questions?

Undergraduate inquiries: chemreg@uga.edu 

Registration and credit transferschemreg@uga.edu

AP Credit, Section Changes, Overrides, Prerequisiteschemreg@uga.edu

Graduate inquiries: chemgrad@uga.edu

Contact Us!

Assistant to the Department Head: Donna Spotts, 706-542-1919 

Main office phone: 706-542-1919 

Main Email: chem-web@franklin.uga.edu

Head of Chemistry: Prof. Jason Locklin