Taxonomic group: fungi / Basidiomycota
(Phylum: Basidiomycota)
Organ / tissue: fruiting body
The structure was elucidated in this paperNCBI PubMed ID: 29352876Publication DOI: 10.1016/j.carbpol.2017.12.017Journal NLM ID: 8307156Publisher: Elsevier
Correspondence: Cheng H <chenghr893
nenu.edu.cn>; Zhou Y <zhouyf383
nenu.edu.cn>
Institutions: Jilin Province Key Laboratory on Chemistry and Biology of Changbai Mountain Natural Drugs, School of Life Sciences, Northeast Normal University, Changchun, China
We isolated and purified a new polysaccharide (WFVP-N-b1) with a molecular weight of 20 kDa from Flammulina velutipes. Results showed that WFVP-N-b1 is composed of an α (1→6)-linked D-galactan backbone and branched at the O-2 of its Galp residues by an α-D-(1→6)-linked Manp attached to t-β-D-Glcp or t-α-D-Fucp side chains. WFVP-N-b1 can significantly induce cytokines secretion and release of toxic molecules. On a cellular level, WFVP-N-b1 is recognized by Toll-like receptor 4 (TLR4). Thereby, the hetero-galactan increased the phosphorylation of mitogen-activated protein kinases (MAPKs) and Akt, promoted degradation of IκB-α and the nuclear translocation of the NF-κB p65 subunit. Importantly, our results indicate that WFVP-N-b1 activated macrophage is mediated by autophagy, as blockade of WFVP-N-b1-induced autophagy by Baf-A1 significantly decreases macrophage activation. This is the first report that hetero-galactan-induced macrophage activation is mediated by autophagy. Collectively, WFVP-N-b1 activated RAW264.7 cells through MAPKs, autophagy, and Akt/NF-κB signaling pathways via TLR4 receptor.
macrophage, Flammulina velutipes, MAPKs, Akt/NF-κB, autophagy, hetero-galactan
Structure type: structural motif or average structure
Location inside paper: p. 212, WFVP-N-b1
Contained glycoepitopes: IEDB_130701,IEDB_134624,IEDB_136045,IEDB_136906,IEDB_137472,IEDB_141794,IEDB_142488,IEDB_142489,IEDB_144562,IEDB_144983,IEDB_146664,IEDB_151528,IEDB_152206,IEDB_152214,IEDB_174333,IEDB_190606,IEDB_983930,IEDB_983931,SB_163,SB_192,SB_44,SB_67,SB_7,SB_72,SB_86
Methods: 13C NMR, 1H NMR, ELISA, Western blotting, HPLC, GPC, extraction, statistical analysis, DEAE, immunofluorescence analyses, determination of NO production, cytokine production, cell viability assay, phenol-sulfuric acid assay, HMBC, ethanol precipitation, centrifugation, phagocytosis assay, HSQC, methylatioon analysis
Biological activity: Immunomodulatory actvities: WFVP-N-b1 significantly activated macrophages as indicated by increasing production of NO, ROS, TNF-α, IL-6 and IL-1β, as well as phagocytic uptake; activated macrophages through MAPKs, autophagy and Akt/NF-κB signaling pathways via TLR4 receptor in vitro and in vivo
NCBI Taxonomy refs (TaxIDs): 38945
Show glycosyltransferases
[as TSV]
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
2 34%aDManp 101.34 68.84 69.69 69.52 77.45 69.36
aDGalp 97.67 77.29 69.26 69.69 68.90 66.94
2,6 27%aLFucp 101.36 71.50 71.26 77.25 73.17 15.37
2,6 73%bDGlcp 102.80 72.86 75.42 69.30 74.70 60.80
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
2 34%aDManp 5.06 4.10 4.02 3.85 3.79 3.83-3.97
aDGalp 5.00 3.94 3.83 4.02 4.09 3.93-4.12
2,6 27%aLFucp 5.03 3.79 3.93 3.96 3.73 1.17
2,6 73%bDGlcp 4.48 3.28 3.44 3.43 3.58 3.71-3.85
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6
2 34%aDManp 101.34/5.06 68.84/4.10 69.69/4.02 69.52/3.85 77.45/3.79 69.36/3.83-3.97
aDGalp 97.67/5.00 77.29/3.94 69.26/3.83 69.69/4.02 68.90/4.09 66.94/3.93-4.12
2,6 27%aLFucp 101.36/5.03 71.50/3.79 71.26/3.93 77.25/3.96 73.17/3.73 15.37/1.17
2,6 73%bDGlcp 102.80/4.48 72.86/3.28 75.42/3.44 69.30/3.43 74.70/3.58 60.80/3.71-3.85
1H NMR data:
| Linkage | Residue | H1 | H2 | H3 | H4 | H5 | H6 |
|---|
| 2 | 34%aDManp | 5.06 | 4.10 | 4.02 | 3.85 | 3.79 | 3.83
3.97 |
| | aDGalp | 5.00 | 3.94 | 3.83 | 4.02 | 4.09 | 3.93
4.12 |
| 2,6 | 27%aLFucp | 5.03 | 3.79 | 3.93 | 3.96 | 3.73 | 1.17 |
| 2,6 | 73%bDGlcp | 4.48 | 3.28 | 3.44 | 3.43 | 3.58 | 3.71
3.85 |
|
13C NMR data:
| Linkage | Residue | C1 | C2 | C3 | C4 | C5 | C6 |
|---|
| 2 | 34%aDManp | 101.34 | 68.84 | 69.69 | 69.52 | 77.45 | 69.36 |
| | aDGalp | 97.67 | 77.29 | 69.26 | 69.69 | 68.90 | 66.94 |
| 2,6 | 27%aLFucp | 101.36 | 71.50 | 71.26 | 77.25 | 73.17 | 15.37 |
| 2,6 | 73%bDGlcp | 102.80 | 72.86 | 75.42 | 69.30 | 74.70 | 60.80 |
|
There is only one chemically distinct structure:
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