Precise Structure and Anticoagulant Activity of Fucosylated Glycosaminoglycan from Apostichopus japonicus: Analysis of Its Depolymerized Fragments
Abstract
:1. Introduction
2. Results and Discussion
2.1. Extraction, Isolation, and Purification of AjFG
2.2. Analysis of Physicochemical Properties
2.3. Preparation of Oligosaccharides with Various dp
2.4. NMR and ESI-Q-TOF-MS Analysis
2.5. Analysis of the Anticoagulant Activity
3. Materials and Methods
3.1. Materials and Chemicals
3.2. Extraction, Isolation and Purification of AjFG
3.3. Determination of Physicochemical Properties
3.4. Preparation and Isolation of Oligosaccharides of AjFG
3.5. NMR and ESI-Q-TOF-MS Analysis
3.6. Determination of Anticoagulant Activities In Vitro
3.7. Human Factor XII Activation and Platelet Aggregation Assays
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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δH (ppm) | Coupling Constant in Hz | δC (ppm) | δH (ppm) | Coupling Constant in Hz | δC (ppm) | ||||||
TI | H-1 | 5.00 | J(1,2) = 4.41 | C-1 | 91.7 | TII | H-1 | 5.00 | J(1,2) = 4.41 | C-1 | 91.6 |
H-2 | 3.98 | J(2,3) = 4.52 | C-2 | 86.1 | H-2 | 3.99 | J(2,3) = 4.52 | C-2 | 86.2 | ||
H-3 | 4.59 | J(3,4) = 5.20 | C-3 | 78.8 | H-3 | 4.60 | J(3,4) = 5.06 | C-3 | 78.4 | ||
H-4 | 4.99 | J(4,5) = 5.10 | C-4 | 79.8 | H-4 | 4.99 | J(2,3) = 5.13 | C-4 | 79.7 | ||
H-5 | 4.48 | J(5,6′) = 9.10 | C-5 | 80.4 | H-5 | 4.48 | J(5,6′) = 9.10 | C-5 | 80.3 | ||
H-6 | 4.29 | J(5,6) = 2.50 | C-6 | 70.6 | H-6 | 4.27 | J(5,6) = 2.47 | C-6 | 70.5 | ||
H-6′ | 4.15 | J(6,6′) =11.29 | H-6′ | 4.16 | J(6,6′) = 11.26 | ||||||
UI | H-1 | 4.56 | J(1,2) = 7.74 | C-1 | 103.3 | UII | H-1 | 4.55 | J(1,2) = 7.44 | C-1 | 103.1 |
H-2 | 3.59 | J(2,3) = 8.29 | C-2 | 75.8 | H-2 | 3.59 | J(2,3) = 8.37 | C-2 | 75.8 | ||
H-3 | 3.60 | J(3,4) = 7.55 | C-3 | 84.2 | H-3 | 3.57 | J(3,4) = 7.78 | C-3 | 84.3 | ||
H-4 | 3.60 | J(4,5) = 9.73 | C-4 | 73.1 | H-4 | 3.62 | J(4,5) = 9.38 | C-4 | 73.1 | ||
H-5 | 3.65 | C-5 | 79.6 | H-5 | 3.65 | C-5 | 79.7 | ||||
C-6 | 178.4 | C-6 | 178.4 | ||||||||
I | H-1 | 5.53 | J(1,2) = 3.92 | C-1 | 99.7 | II | H-1 | 5.31 | J(1,2) = 3.98 | C-1 | 101.8 |
H-2 | 4.39 | J(2,3) = 10.58 | C-2 | 77.9 | H-2 | 3.90 | J(2,3) = 10.46 | C-2 | 69.2 | ||
H-3 | 4.10 | J(3,4) = 2.96 | C-3 | 69.2 | H-3 | 4.57 | J(3,4) = 2.95 | C-3 | 78.1 | ||
H-4 | 4.62 | C-4 | 83.7 | H-4 | 4.83 | C-4 | 81.9 | ||||
H-5 | 4.50 | J(5,6) = 6.80 | C-5 | 68.9 | H-5 | 4.46 | J(5,6) = 6.62 | C-5 | 69.2 | ||
H-6 | 1.19 | C-6 | 18.4 | H-6 | 1.19 | C-6 | 18.6 |
Sample | Mw (kDa) | APTT (μg/mL) a | Anti-FXase (IC50, ng/mL) b |
---|---|---|---|
AjFG | 76.4 | 3.06 | 9.20 |
dAjFG | 3.90 | 26.5 | 189 |
4 | 3.49 | 10.3 | 131 |
3 | 2.63 | 20.1 | 200 |
2 | 1.77 | >128 | >1000 |
1 | 0.93 | >128 | >1000 |
LMWH | 4.50 | 11.3 | 128 |
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Guan, R.; Peng, Y.; Zhou, L.; Zheng, W.; Liu, X.; Wang, P.; Yuan, Q.; Gao, N.; Zhao, L.; Zhao, J. Precise Structure and Anticoagulant Activity of Fucosylated Glycosaminoglycan from Apostichopus japonicus: Analysis of Its Depolymerized Fragments. Mar. Drugs 2019, 17, 195. https://doi.org/10.3390/md17040195
Guan R, Peng Y, Zhou L, Zheng W, Liu X, Wang P, Yuan Q, Gao N, Zhao L, Zhao J. Precise Structure and Anticoagulant Activity of Fucosylated Glycosaminoglycan from Apostichopus japonicus: Analysis of Its Depolymerized Fragments. Marine Drugs. 2019; 17(4):195. https://doi.org/10.3390/md17040195
Chicago/Turabian StyleGuan, Ruowei, Yuan Peng, Lutan Zhou, Wenqi Zheng, Xixi Liu, Pin Wang, Qingxia Yuan, Na Gao, Longyan Zhao, and Jinhua Zhao. 2019. "Precise Structure and Anticoagulant Activity of Fucosylated Glycosaminoglycan from Apostichopus japonicus: Analysis of Its Depolymerized Fragments" Marine Drugs 17, no. 4: 195. https://doi.org/10.3390/md17040195