The impact of structural modification of sulfated polysaccharides on bone morphogenic protein 2 and inhibition of endothelial cell angiogenesis.

Sulfated polysaccharides play important roles in angiogenesis. However, the impact of structural alteration of sulfated polysaccharide on the bioactivity is still vague. In this study, binding between different sulfated polysaccharides and bone morphogenic protein 2 (BMP2) was measured to understand the sense of this motif transformation. The results showed that binding between sulfated α-1,4-glucan and BMP2 was the most intensive. The branch of α-1,4-glucan was important for the binding. The affinity of sulfated polysaccharides to BMP2 increased as the molecular weight (MW) and degree of substitution (DS) increased. DS that exceeded 1.05 impaired binding and played more important role in polysaccharide BMP2 interaction than MW. The reservation of partial 6-OH would benefit its binding ability to BMP2. Further, we showed that sulfated polysaccharides with strong binding to BMP2 blocked phosphorylation of Smad 1/5/8 and expression of Id1 to a greater extent than those not strongly bind to BMP2. The binding strength of polysaccharides to BMP2 increased, so did the potency of the anti-angiogenesis effects.

Semi-volatile organic compounds in tap water from Hangzhou, China: Influence of pipe material and implication for human exposure.

Investigations on environmental behaviors of SVOCs have recently received great attention. However, the SVOC occurrence and influence of pipe materials on SVOC levels in the tap water have received little attention. Herein, we collected tap water samples from 25 households constructed at different ages in Hangzhou, China. Concentrations of 61 SVOCs, including phthalate esters (PAEs), organochlorine pesticides (OCPs), polycyclic aromatic hydrocarbons (PAHs), and polychlorinated biphenyls (PCBs), were simultaneously determined in these collected samples. The potential human exposure risks were evaluated via the hazard index calculation. Our results showed that the total concentration of detected SVOCs (∑SVOCs) ranged from 110 to 289 µg/L (mean, 179 µg/L), and the SVOCs were dominated by PAHs (mean, 116 ng/L) and PAEs (mean, 55 ng/L) in Hangzhou tap water. 12 PCB congeners were detected in Hangzhou tap water samples, with hepta-CBs (68% of samples) as the most frequently detected PCBs. Nearly all tap water samples contained measurable o, p'-DDE, p, p'-DDT, and p, p'-DDD, and ∑DDTs had significantly (p < 0.05) higher concentrations than ∑HCHs. All target PAHs had high detection frequencies (>72%) in tap water samples, with their mean concentrations in the range of 2.1-41 ng/L. Tap water from steel pipes had relatively lower SVOC concentrations than that from either reinforced concrete, gray cast iron, or ductile iron pipes. Although no carcinogenic risks originating from exposure to SVOCs through ingestion and bathing were observed, the tap water from steel pipes showed relatively low exposure risks than that from other materials. Data provided here, for the first, are helpful in understanding the influence of pipe materials on human SVOC exposure risks through tap water intake.

A fucoidan from Nemacystus decipiens disrupts angiogenesis through targeting bone morphogenetic protein 4.

A sulfated and acetylated fucoidan, named NDH01, was extracted from seaweed Nemacystus decipiens. NDH01 was composed of mannose, glucuronic acid, fucose, sulfate group and acetyl group in the molar ratio of 3.0: 14.4: 82.6: 34.3: 13.9. The backbone of NDH01 was fucose-free core, composed of α-d-1,2-Manp and ß-d-1,4-GlcpA disaccharide repeat unit. The branches were attached at the C3, C4 and C6 of α-d-1,2-Manp. The sidechain was composed of α-l-1,3,4-Fucp, α-l-1,4-Fucp, α-l-1,3-Fucp and α-l-1,4-GlcpA. The sulfate group was linked to C4 of α-l-1,3,4-Fucp, whereas, acetyl group was branched on C2 of α-l-1,2,3-Fucp. NDH01 could disrupt tube formation and inhibit the migration as well as cell growth of human microvascular endothelial cells. Besides, phosphorylation of Smad/1/5/8, Erk and FAK was significantly inhibited by NDH01. Further studies uncovered that NDH01 blocked Smad1/5/8 signaling via interacting with bone morphogenetic protein 4 and downregulating bone morphogenetic protein 4 expression. The results suggested that NDH01 might be an angiogenesis inhibitor through targeting bone morphogenetic protein 4.

Structural elucidation of a pectin from flowers of Lonicera japonica and its antipancreatic cancer activity.

To investigate polysaccharide structure from Lonicera japonica, and study its effects on behavior of pancreatic cells, a homogenous polysaccharide, LJ-02-1, was extracted and purified from flowers of L. japonica by DEAE-cellulose and Sephacryl S-200HR column. The molecular weight was estimated to be 54kDa. Monosaccharide composition was determined to be rhamnose, galacturonic acid, galactose and arabinose in the molar ratio of 10.77:7.88:15.45:65.89 by analyzing the PMP derivatives of the monosaccharides from 2M trifluoracetic acid hydrolysis via HPLC. Based on methylation analysis, partial acid hydrolysis, and NMR spectra, the polysaccharide was elucidated to be a rhamnogalacturonan backbone and substituted partly at C-4 of rhamnose. The branches were determined to be T- and 1,4,6-linked ß-d-Galp, T- and 1,5-linked α-l-Araf. The polysaccharide might inhibit BxPC-3 and PANC-1 pancreatic cancer cells growth at the concentration of 1mg/mL with inhibitory ratio of 66.7% and 52.1%, respectively.

Structural and functional study of D-glucuronyl C5-epimerase.

Heparan sulfate (HS) is a glycosaminoglycan present on the cell surface and in the extracellular matrix, which interacts with diverse signal molecules and is essential for many physiological processes including embryonic development, cell growth, inflammation, and blood coagulation. D-glucuronyl C5-epimerase (Glce) is a crucial enzyme in HS synthesis, converting D-glucuronic acid to L-iduronic acid to increase HS flexibility. This modification of HS is important for protein ligand recognition. We have determined the crystal structures of Glce in apo-form (unliganded) and in complex with heparin hexasaccharide (product of Glce following O-sulfation), both in a stable dimer conformation. A Glce dimer contains two catalytic sites, each at a positively charged cleft in C-terminal α-helical domains binding one negatively charged hexasaccharide. Based on the structural and mutagenesis studies, three tyrosine residues, Tyr(468), Tyr(528), and Tyr(546), in the active site were found to be crucial for the enzymatic activity. The complex structure also reveals the mechanism of product inhibition (i.e. 2-O- and 6-O-sulfation of HS keeps the C5 carbon of L-iduronic acid away from the active-site tyrosine residues). Our structural and functional data advance understanding of the key modification in HS biosynthesis.