A further unique chondroitin sulfate from the shrimp Litopenaeus vannamei with antithrombin activity that modulates acute inflammation

Abstract

The detailed structure of a further Chondroitin Sulfate from Litopenaeus vannamei shrimp (sCS) is described. The backbone structure was established by 1 H/ 13 C NMR, which identified 3-O-sulfated GlcA, 4-O-sulfated GalNAc, 6-O-sulfated GalNAc, and 4,6-di-O-sulfated GalNAc residues. GlcA is linked to GalNAc 4,6 di S and GlcA 3S is linked to GalNAc 4S, GalNAc 4,6 di-S and GalNAc6S residues. The anticoagulant properties of this sCS were evaluated by activated partial thromboplastin time, anti-IIa, anti-Xa and anti-heparin cofactor II-mediated activities, and sCS failed to stabilise antithrombin in a fluoresence shift assay. The anti-inflammatory effect of sCS was explored using a model of acute peritonitis, followed by leukocyte count and measurement of the cytokines, IL-1β, IL-6 and TNF-α. The compound showed low clotting effects, but high anti-IIa activity and HCII-mediated thrombin inhibition. Its anti-inflammatory effect was shown by leukocyte recruitment inhibition and a decrease in pro-inflammatory cytokine levels. Although the biological role of sCS remains unknown, its properties indicate that it is suitable for studies of multi-potent molecules obtained from natural sources.

NMR Characterization of the Interactions Between Glycosaminoglycans and Proteins

“…Oversulfated chondroitin sulfate (OSCS) was sulfated at all sites that could be sulfated, and it was one of the culprits that triggered the "heparin crisis" in 2008 (Zhu et al, 2019). There is a special kind of 3-O-sulfated chondroitin sulfate (CS-K) in marine organisms that has a high affinity for growth factors (Palhares et al, 2019).…”

Isolation and Characterization of a Heparin-Like Compound with Potent Anticoagulant and Fibrinolytic Activity from the Clam Coelomactra antiquata

“…The structure of G15 was further analyzed by two-dimensional NMR techniques of COSY, TOCSY, NOESY, and HSQC, and its chemical shifts were assigned based on these spectrograms and previous reports (Table 1) [27][28][29][30][31]. In general, when no group substitution occurs, the signals of C2, C3, and C4 were between 70 and 77 ppm, and the signals of C6 were between 60 and 64 ppm.…”

“…The NMR signals attributed to the N,3,6-trisulfated glucosamine residue, a typical marker of the pentasaccharide sequence of the active site of heparin and heparan sulfates for antithrombin binding, were detected in the spectra of the clam heparin-like compound, indicating that the anticoagulant activity of such compounds may be comparable to that of heparin. The structure of G15 was further analyzed by two-dimensional NMR techniques of COSY, TOCSY, NOESY, and HSQC, and its chemical shifts were assigned based on these spectrograms and previous reports (Table 1) [27][28][29][30][31]. In general, when no group substitution occurs, the signals of C2, C3, and C4 were between 70 and 77 ppm, and the signals of C6 were between 60 and 64 ppm.…”

Chondroitin sulfate from fish waste exhibits strong intracellular antioxidant potential

“…For this reason, there is great interest in expanding studies on the role of CS in these physiological events. In this work, an extraction procedure similar to those used previously to obtain compounds with pharmacological potential in different invertebrates and marine species was performed, showing efficiency and reproducibility ( 11 , 17 ). The characterized CS-A tilapia , containing 59.6% disaccharides sulfated at position 4 of galactosamine, has been described for the first time.…”