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

Abstract

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.

The Landscape of Small Leucine-Rich Proteoglycan Impact on Cancer Pathogenesis with a Focus on Biglycan and Lumican

“…Thus, bisulfated saturated and trisulfated unsaturated disaccharides were detected. IMS MS/MS analysis demonstrated that at the CS/DS chains, non-reducing end 3-O-sulfated GlcA harbors a rare bisulfated motif that is possibly relevant to cancerassociated signaling [20]. In addition, biglycan can bear N-linked oligosaccharide chains bound into its central LRRs motifs, which are able to specifically modify biglycan-binding abilities [59].…”

“…SLRPs interact with structural ECM molecules, cell membrane receptors, growth factors, and cytokines, as well as with their protein cores and GAG chains [16,17]. Notably, GAG chains can modify protein core-ligand interactions by introducing immense variability in their signaling [18][19][20]. A plethora of studies have demonstrated that SLRPs' expression is altered in tumors [21,22] and that by interacting with tyrosine kinase receptors or growth factors, they affect tumor behavior [23][24][25][26].…”

FAIMS Shotgun Lipidomics for Enhanced HILIC-like Separation and Automated Annotation of Gangliosides

“…Several devices with different combinations of the applied electric field and separation technique are commercially available 21,22 . Numerous studies using IMS based on temporal-separation have been reported for the analysis of gangliosides, offering separation of isomeric ganglioside species and deep characterization of the ganglioside lipidome 16,17,[23][24][25][26][27][28][29][30][31][32][33][34] . In contrast, spatialseparation based IMS for gangliosides analysis has received limited attention 35 , despite its potential for increased sensitivity as shown in proteomics and lipidomics studies [36][37][38][39][40][41][42][43][44] .…”

FAIMS Shotgun Lipidomics for Enhanced Class- and Charge-State Separation Complemented by Automated Ganglioside Annotation

“…Numerous studies using IMS based on temporal-separation have been reported for the analysis of gangliosides, offering separation of isomeric ganglioside species and deep characterization of the ganglioside lipidome. ,,− In contrast, spatial-separation based IMS for gangliosides analysis has received limited attention, despite its potential for increased sensitivity as shown in proteomics and lipidomics studies. − Overall, ganglioside analysis approaches aim for high-confidence identifications, requiring accurate mass, MS/MS fragment information, and, ideally, matching database entries or a reference standard . Collision-induced dissociation (CID) and high-energy collisional dissociation (HCD) are the most commonly used ion activation methods for MS/MS lipid structural analysis.…”