Orbitrap mass spectrometry characterization of hybrid chondroitin/dermatan sulfate hexasaccharide domains expressed in brain

Robu, Adrian; Popescu, Laurentiu; Munteanu, Cristian V. A.; Seidler, Daniela G.; Zamfir, Alina D.

doi:10.1016/j.ab.2015.06.028

Cited by 9 publications

(11 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Different studies have reported the comprehensive MS characterization of the various sulfated dp2 isomers from glucosaminoglycans allowing decisive progresses in the compositional analysis and profiling of heparin and heparan sulfate from multiple origins . Less data are comparatively available for galactosaminoglycan‐derived oligosaccharides, especially as regards the detailed characterization of chondroitin sulfate oligosaccharides . Previous studies showed that MS/MS fragmentation of CS‐A and CS‐C dp2 produced identical Y 1 and Z 1 fragments with different respective intensities .…”

Section: Introductionmentioning

confidence: 99%

Isomer separation and effect of the degree of polymerization on the gas‐phase structure of chondroitin sulfate oligosaccharides analyzed by ion mobility and tandem mass spectrometry

Poyer

Lopin‐Bon

Jacquinet

et al. 2017

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

show abstract

Section: Introductionmentioning

confidence: 99%

Isomer separation and effect of the degree of polymerization on the gas‐phase structure of chondroitin sulfate oligosaccharides analyzed by ion mobility and tandem mass spectrometry

Poyer

Lopin‐Bon

Jacquinet

et al. 2017

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

show abstract

“…The ion assignment to species containing 4,5‐Δ‐GlcA was based on the mechanism of chondroitin AC lyase cleavage reaction, forming a 4,5 double bond in the GlcA residue. According to previous observations, chondroitin AC lyase cleaves the 3GalNAcβ1‐4GlcAβ1 glycosidic bond, irrespective of the sulfation content, distribution along the chain and/or the site(s) in the oligosaccharide ring.…”

Section: Resultsmentioning

confidence: 62%

“…Prior to the CS extraction and purification procedures, which followed the protocol previously described by us, the HEK293 cells were transfected with human decorin cDNA as shown before . Briefly, the sample was centrifuged, and the supernatant was dissolved in Tris/HCl buffer (pH 7.4) containing 150 mM NaCl and applied on a DEAE‐anion exchange column (Pall Life Sciences, Dreieich, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…For this reason, the NanoMate robot encompassing automatic sample infusion using nanoESI was coupled to different types of instruments for the identification and structural characterization of CS/DS, such as the high‐capacity ion trap (HCT) or the quadrupole time‐of‐flight (QTOF) . This method, together with a fragmentation technique such as collision‐induced dissociation tandem MS (CID MS/MS) proved its feasibility in discovering and structural characterization of novel molecular species with a potential biomarker role in various biological matrices as shown by our previous work . Additionally, in view of the multiple benefits of chip‐based analyses, the progress of the microfluidic and nanofluidic systems for MS come into sight as an important research topic—some of the advantages of this method being represented by the possibility to map and sequence various classes of biomolecules in a high‐throughput manner and the outstanding performance in terms of sensitivity, reproducibility, and data quality …”

Section: Introductionmentioning

confidence: 97%

“…The disaccharide part is modified at the N ‐acetylgalactosamine (GalNAc) moiety by sulfation. GalNAc unit might be sulfated in position 4 or 6, while, if a supplementary sulfate group is present, in some oversulfated species, a small amount of the uronic acid might be sulfated in position 2 . The biological properties of CS/DS are thus influenced by the degree of sulfation, which does not occur randomly, and also, by the distribution of the sulfate group in the disaccharide moiety …”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Chip‐based high resolution tandem mass spectrometric determination of fibroblast growth factor—chondroitin sulfate disaccharides noncovalent interaction

et al. 2018

Self Cite

View full text Add to dashboard Cite

Fibroblast growth factor-2 (FGF-2) is involved in wound healing and embryonic development. Glycosaminoglycans (GAGs), the major components of the extracellular matrix (ECM), play fundamental roles at this level. FGF-GAG noncovalent interactions are in the focus of research, due to their influence upon cell proliferation and tissue regeneration. Lately, high resolution mass spectrometry (MS) coupled with chip-nanoelectrospray (nanoESI) contributed a significant progress in glycosaminoglycomics by discoveries related to novel species and their characterization. We have employed a fully automated chip-nanoESI coupled to a quadrupole time-of-flight (QTOF) MS for assessing FGF-GAG noncovalent complexes. For the first time, a CS disaccharide was involved in a binding assay with FGF-2. The experiments were conducted in 10 mM ammonium acetate/formic acid, pH 6.8, by incubating FGF-2 and CS in buffer. The detected complexes were characterized by top-down in tandem MS (MS/MS) using collision induced-dissociation (CID). CID MS/MS provided data showing for the first time that the binding process occurs via the sulfate group located at C4 in GalNAc. This study has demonstrated that chip-MS may generate reliable data upon the formation of GAG-protein complexes and their structure. Biologically, the findings are relevant for studies focused on the identification of the active domains in longer GAG chains.

show abstract

Glycomics by ion mobility tandem mass spectrometry of chondroitin sulfate disaccharide domain in biglycan

et al. 2023

Self Cite

View full text Add to dashboard Cite

Biglycan (BGN), a small leucine-rich repeat proteoglycan, is involved in a variety of pathological processes including malignant transformation, for which the upregulation of BGN was found related to cancer cell invasiveness. Because the functions of BGN are mediated by its chondroitin/dermatan sulfate (CS/DS) chains through the sulfates, the determination of CS/DS structure and sulfation pattern is of major importance. In this study, we have implemented an advanced glycomics method based on ion mobility separation (IMS) mass spectrometry (MS) and tandem MS (MS/MS) to characterize the CS disaccharide domains in BGN. The high separation efficiency and sensitivity of this technique allowed the discrimination of five distinct CS disaccharide motifs, of which four irregulated in their sulfation pattern. For the first time, trisulfated unsaturated and bisulfated saturated disaccharides were found in BGN, the latter species documenting the non-reducing end of the chains. The structural investigation by IMS MS/MS disclosed that in one or both of the CS/DS chains, the non-reducing end is 3-O-sulfated GlcA in a rather rare bisulfated motif having the structure 3-O-sulfated GlcA-4-O-sulfated GalNAc. Considering the role played by BGN in cancer cell spreading, the influence on this process of the newly identified sequences will be investigated in the future.

show abstract

Orbitrap mass spectrometry characterization of hybrid chondroitin/dermatan sulfate hexasaccharide domains expressed in brain

Cited by 9 publications

References 35 publications

Isomer separation and effect of the degree of polymerization on the gas‐phase structure of chondroitin sulfate oligosaccharides analyzed by ion mobility and tandem mass spectrometry

Isomer separation and effect of the degree of polymerization on the gas‐phase structure of chondroitin sulfate oligosaccharides analyzed by ion mobility and tandem mass spectrometry

Chip‐based high resolution tandem mass spectrometric determination of fibroblast growth factor—chondroitin sulfate disaccharides noncovalent interaction

Glycomics by ion mobility tandem mass spectrometry of chondroitin sulfate disaccharide domain in biglycan

Contact Info

Product

Resources

About