Precise Structure and Anticoagulant Activity of Fucosylated Glycosaminoglycan from Apostichopus japonicus: Analysis of Its Depolymerized Fragments

Abstract

Apostichopus japonicus is one of the most economically important species in sea cucumber aquaculture in China. Fucosylated glycosaminoglycan from A. japonicus (AjFG) has shown multiple pharmacological activities. However, results from studies on the structure of AjFG are still controversial. In this study, the deaminative depolymerization method that is glycosidic bond-selective was used to prepare the depolymerized products from AjFG (dAjFG), and then a series of purified oligosaccharide fragments such as tri-, hexa-, nona-, and dodecasaccharides were obtained from dAjFG by gel permeation chromatography. The 1D/2D NMR and ESI-MS spectrometry analyses showed that these oligosaccharides had the structural formula of l-FucS-α1,3-d-GlcA-β1,3-{d-GalNAc4S6S-β1,4-[l-FucS-α1,3-]d-GlcA-β1,3-}n-d-anTal-diol4S6S (n = 0, 1, 2, 3; FucS represents Fuc2S4S, Fuc3S4S, or Fuc4S). Thus, the unambiguous structure of native AjFG can be rationally deduced: it had the backbone of {-4-d-GlcA-β1,3-d-GalNAc4S6S-β1-}n, which is similar to chondroitin sulfate E, and each d-GlcA residue in the backbone was branched with a l-FucS monosaccharide at O-3. Bioactivity assays confirmed that dAjFG and nonasaccharides and dodecasaccharides from AjFG had potent anticoagulant activity by intrinsic FXase inhibition while avoiding side effects such as FXII activation and platelet aggregation.

Structural Characterization and Heparanase Inhibitory Activity of Fucosylated Glycosaminoglycan from Holothuria floridana

“…The signals at 4.98 ppm and 4.95 ppm could be assigned to H-1 and H-4 of the T residue, according to the literature [22,32], indicating that the new anTal residue at the reducing end was formed. Six distinct signals were observed at the region of 5.2-5.7 ppm, which should be the anomeric protons of fucosyl residues varying in pattern of sulfation, as previously described [22]. According to the 1 H-1 H COSY spectrum (Figure 4a), the other H signals of the resonance systems could be further assessed ( Table 1).…”

“…It has been used to prepare the depolymerized products of FG from S. variegatus and A. japonicus , etc. [ 22 , 32 ] These reports on the structure analysis of the depolymerization products revealed that the procedure only breaks the backbone glycosidic bonds and has little effect on other structures of native FG. In the present study, dHfFG was prepared from the native HfFG by deacetylated-deaminative depolymerization with the yields of 60.0%.…”

“…The development of chemoselective depolymerization methods could facilitate the progress in the structural analysis of FG from various species. Recently, the deaminative degradation [ 21 , 22 ] and β-eliminative depolymerization methods were used to investigate the exact structure and the structure-activity relationship of FG, and these strategies have been used in FG from Stichopus japonicus [ 22 ], Stichopus horrens [ 23 ], Bohadschia argus , Holothuria coluber [ 24 ], Actinopyga miliaris , Holothuria albiventer and Stichopus variegatus [ 25 ]. The structure exploration of FG through the bottom-up strategy based on the structural analysis of purified oligosaccharides from chemoselective depolymerization methods has proved useful in confirming the structure of the CS-like core and sulfated fucose branches.…”

Collective Locomotion of Human Cells, Wound Healing and Their Control by Extracts and Isolated Compounds from Marine Invertebrates

“…It is widely recognized that most bioactive properties of sea cucumbers derive from components extracted from their body wall. For example, in the case of S. japonicus total extracts exhibited anti-oxidant characteristics, and isolated molecules, such as unsaturated fatty acids and a fucosylated chondroitin sulfate glycosaminoglycan, were proven to exert α-glucosidase inhibitory, immunostimulatory, anti-coagulant, and hypolipidemic activities [180][181][182][183][184]. Collagen, which is the major component of the body wall of S. japonicus [185], has been studied for its potential as an additive and functional food ingredient that is capable of maintaining food quality thanks to its gelling properties [186,187].…”

Marine Antithrombotics

“…Depolymerized fragments of this glycan also exhibited an aPTT prolongation and anti-FXase activity, although the effect was smaller than that of native sugar, signifying the effect of chain shortening on the function. The requirement of a nonasaccharide, in terms of chain length, appeared to be necessary for the anticoagulant activity of A. japonicas FucCS [ 66 ]. Fucosylated glycan (FCS hm ) reported from Holothuria mexicana was found to bear a unique fucosyl branch attached to the O6 of GalNAc, besides the regular fucosyl unit linked to the O3 position of GlcA.…”