Structural Characterization and Heparanase Inhibitory Activity of Fucosylated Glycosaminoglycan from Holothuria floridana
Abstract
:1. Introduction
2. Results and Discussion
2.1. Extraction, Isolation and Purification of HfFG
2.2. Chemical Compositions and Physicochemical Properties of HfFG
2.3. Preparation and Structure Characteristics of the Low-Molecular-Weight HfFG (dHfFG)
2.4. Purification of Homogeneous Oligosaccharides Fragments
2.5. NMR Analysis of dHfFG, Fr-1, Fr-2 and Fr-3
2.6. Heparanase Inhibitory Activity
3. Materials and Methods
3.1. Materials
3.2. Extraction, Isolation and Purification of Polysaccharides
3.3. Preparation of Low-Molecular-Weight HfFG by the Deacetylation–Deaminative Cleavage Method
3.4. Purification of Homogeneous Oligosaccharide Fragments from dHfFG
3.5. General Procedures for Physicochemical Properties Determination
3.6. NMR Analysis
3.7. Heparanase Inhibitory Activities Assay
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Residue. | Chemical Shift (ppm) | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
H/C | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | ||
T | →3)-β-d-anTal4S6S-diol | H | 4.983 | 3.949 | 4.523 | 4.952 | 4.433 | 4.263/4.119 | ||
C | 91.58 | 85.93 | 79.13 | 79.79 | 80.38 | 70.6 | ||||
U’ | →4)-β-d-GlcA-(1→ | H | 4.484 | 3.623 | 3.673 | 3.893 | 3.673 | |||
C | 103.64 | 76.38 | 79.79 | 78.13 | 79.32 | 177.34 | ||||
A | →3)-β-d-GalNAc4S6S-(1→ | H | 4.485 | 3.971 | 3.938 | 4.729 | 3.893 | 4.101/4.184 | ||
C | 102.58 | 54.05 | 78.9 | 79.05 | 74.53 | 69.88 | 177.76 | 25.3 | ||
F’ (I’) | α-l-Fuc2S4S-(1→ | H | 5.602 | 4.403 | 4.064 | 4.766 | 4.817 | 1.276 | ||
C | 99.3 | 77.88 | 67.27 | 83.62 | 69.04 | 18.53 | ||||
F’ (II’) | α-l-Fuc3S4S-(1→ | H | 5.261 | 3.857 | 4.422 | 4.942 | 4.783 | 1.303 | ||
C | 101.85 | 69.18 | 77.75 | 81.92 | 68.89 | 18.8 | ||||
F’(III’) | α-l-Fuc4S-(1→ | H | 5.315 | 3.708 | 3.949 | 4.69 | 4.745 | 1.267 | ||
C | 102.04 | 71.33 | 71.46 | 83.94 | 68.98 | 18.47 | ||||
U | d-GlcA-(1→ | H | 4.416 | 3.512 | 3.551 | 3.618 | 3.671 | |||
C | 106.4 | 76.63 | 84.19 | 72.6 | 79.32 | 177.76 | ||||
F (I) | α-l-Fuc2S4S-(1→ | H | 5.493 | 4.357 | 4.079 | 4.596 | 4.452 | 1.167 | ||
C | 99.17 | 77.88 | 69.18 | 83.83 | 68.89 | 18.35 | ||||
F (II) | α-l-Fuc3S4S-(1→ | H | 5.273 | 3.856 | 4.437 | 4.933 | 4.769 | 1.167 | ||
C | 101.55 | 69.1 | 78.06 | 82.13 | 69.1 | 18.5 | ||||
F (III) | α-l-Fuc4S-(1→ | H | 5.187 | 3.711 | 3.93 | 4.511 | 4.408 | 1.144 | ||
C | 101.19 | 71.33 | 71.46 | 83.55 | 68.98 | 18.35 |
Sample | Mw (Da) | Specific Rotation [α] | IC50 (ng/mL) | IC50 (nM) |
---|---|---|---|---|
HfFG | 47,282 | −50° | 11.4 ± 1.39 | 0.24 ± 2.94 |
Fr-3 | 3652 a | −45° | 2346 ± 409 | 642.3 ± 112 |
Fr-2 | 2767 | −32° | 2766 ± 381 | 999.4 ± 138 |
Fr-1 | 1841 | −44° | 4496 ± 590 | 2440 ± 320 |
Heparin | ~18,000 | -- | 32.3 ± 2.21 | 1.79 ± 0.12 |
LMWH | ~4500 | -- | 300 ± 29.4 | 66.67 ± 6.53 |
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Shi, X.; Guan, R.; Zhou, L.; Zuo, Z.; Tao, X.; Wang, P.; Zhou, Y.; Yin, R.; Zhao, L.; Gao, N.; et al. Structural Characterization and Heparanase Inhibitory Activity of Fucosylated Glycosaminoglycan from Holothuria floridana. Mar. Drugs 2021, 19, 162. https://doi.org/10.3390/md19030162
Shi X, Guan R, Zhou L, Zuo Z, Tao X, Wang P, Zhou Y, Yin R, Zhao L, Gao N, et al. Structural Characterization and Heparanase Inhibitory Activity of Fucosylated Glycosaminoglycan from Holothuria floridana. Marine Drugs. 2021; 19(3):162. https://doi.org/10.3390/md19030162
Chicago/Turabian StyleShi, Xiang, Ruowei Guan, Lutan Zhou, Zhichuang Zuo, Xuelin Tao, Pin Wang, Yanrong Zhou, Ronghua Yin, Longyan Zhao, Na Gao, and et al. 2021. "Structural Characterization and Heparanase Inhibitory Activity of Fucosylated Glycosaminoglycan from Holothuria floridana" Marine Drugs 19, no. 3: 162. https://doi.org/10.3390/md19030162