Among Dissociation Methods, Negative Electron Transfer Dissociation (NETD) Has Been Proven the Most Useful for Glycosaminoglycan (GAG) Sequencing Because It Produces Informative Fragmentation, a Low Degree of Sulfate Losses, High Sensitivity, and Translatability to Multiple Instrument Types. the Challenge, However, is to Distinguish Positional Sulfation. in Particular, NETD Has Been Reported to Fail to Differentiate 4-O- Versus 6-O-Sulfation in Chondroitin Sulfate Decasaccharide. This Raised the Concern of Whether NETD is Able to Differentiate the Rare 3-O-Sulfation from Predominant 6-O-Sulfation in Heparan Sulfate (HS) Oligosaccharides. Here, We Report that NETD Generates Highly Informative Spectra that Differentiate Sites of O-Sulfation on Glucosamine Residues, Enabling Structural Characterizations of Synthetic HS Isomers Containing 3-O-Sulfation. Further, Lyase-Resistant 3-O-Sulfated Tetrasaccharides from Natural Sources Were Successfully Sequenced. Notably, for All of the Oligosaccharides in This Study, the Successful Sequencing is based on NETD Tandem Mass Spectra of Commonly Observed Deprotonated Precursor Ions Without Derivatization or Metal Cation Adduction, Simplifying the Experimental Workflow and Data Interpretation. These Results Demonstrate the Potential of NETD as a Sensitive Analytical Tool for Detailed, High-Throughput Structural Analysis of Highly Sulfated GAGs. [Figure Not Available: See Fulltext.].


Chemical and Biochemical Engineering


National Institutes of Health, Grant R21HL131554

Keywords and Phrases

Fourier transform ion cyclotron resonance mass spectrometry; Glycomics; Glycosaminoglycan; Heparan sulfate; Negative electron transfer dissociation; Sulfation

International Standard Serial Number (ISSN)

1879-1123; 1044-0305

Document Type

Article - Journal

Document Version


File Type





© 2023 American Chemical Society, All rights reserved.

Publication Date

01 Jun 2018

PubMed ID