bims-supasi 2022-01-23 papers

Mol Cell Endocrinol. 2022 Jan 15. pii: S0303-7207(22)00008-9. [Epub ahead of print] 111561

Bisphenol A-sulfate conjugate disrupts AURKA transcription and cell cycle in BeWo cytotrophoblasts.

Jumpei Fujiki, Megumi Uchida, Sakurako Tsunoda, Naoyuki Maeda, Hiroki Inoue, Hiroshi Yokota, Hidetomo Iwano.

Bisphenol A (BPA) has been shown to exhibit various toxic effects, including the induction of reproductive disorders. Generally, BPA is converted to conjugated metabolites, leading to bio-inactivation. On the other hand, the toxicity of conjugated metabolites is not fully understood. Notably, the placenta develops the sulfate-sulfatase pathway, which transports and reactivates sulfated steroids. Therefore, we investigated the potential adverse effects of the BPA-sulfate conjugate (BPA-S) on human placenta-derived BeWo cytotrophoblasts. In the present study, high-concentration BPA-S (100 μM) induced significant inhibition of BeWo growth, with effects similar to those seen with unconjugated BPA (100 μM and 100 nM). This growth inhibition was restored by treatment of the cells with an inhibitor of the organic anion-transporting peptides (OATPs) (bromosulphophthalein) or with a sulfatase (STS) inhibitor (STX64). BeWo exhibits expression of the genes encoding OATP1A2 and OATP4A1 as known sulfated steroid transporters and STS, suggesting that BPA-S suppresses cell growth activity via the sulfate-sulfatase pathway. In addition, cell cycle analysis revealed that BPA-S (100 μM) increased the fraction of cytotrophoblasts in the G2/M phases and significantly decreased the accumulation of the transcript encoding Aurora kinase A (AURKA), which is a critical regulator of cellular division. These results suggested that BPA-S triggers cell cycle arrest and inhibits proliferation of BeWo cytotrophoblasts by decreased AURKA, an effect that is mediated by the sulfate-sulfatase pathway. Overall, these findings provide insights into the reactivation of sulfated endocrine-disrupting chemicals and subsequent adverse effects.

Keywords: BPA; Bisphenol; Endocrine-disrupting chemicals (EDCs); Placenta; Sulfate-sulfatase pathway

DOI: https://doi.org/10.1016/j.mce.2022.111561

Food Funct. 2022 Jan 20.

Heteroconjugates of quercetin with 4'-O-sulfate selectively accumulate in rat plasma due to limited urinary excretion.

Seiya Tanaka, Aphichat Trakooncharoenvit, Miyu Nishikawa, Shinichi Ikushiro, Hiroshi Hara.

Quercetin and methylquercetin are present in a variety of sulfate and glucuronide conjugates in the plasma of quercetin-fed rats and humans. Quercetin conjugates exhibit various physiological activities, depending on the type and position of conjugation. However, little is known regarding the type and position of isomers of quercetin conjugates in the plasma, their accumulation in the liver and kidneys, and their excretion via urine. Using authentic standards of quercetin conjugates and liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis, we identified and quantified various quercetin conjugates in blood plasma, urine, liver, and kidney tissues 1, 4, and 10 h after orally administering 33.1 μmol kg-1 quercetin glucosides to rats. The profiles of quercetin conjugates were largely different among plasma, urine, liver, and kidneys. Very limited heteroconjugates (7-O-glucuronide-4'-O-sulfate) of quercetin and methylquercetin dominated in the plasma, but these heteroconjugates were much less excreted via urine and did not largely accumulate in the liver and kidneys. Heteroconjugates constituting sulfates other than 4' position sulfate, 7-O-glucuronide-3'-O-sulfate, 4'-O-glucuronide-7-O-sulfate, and 3'-O-glucuronide-7-O-sulfate were major metabolites in urine, but were minimally detected in the plasma. We also found that mono-sulfate conjugates were abundant in the liver and renal tissues. These results suggest that excretion of quercetin conjugates, especially heteroconjugates, into urine is highly selective. The heteroconjugates with 4'-O-sulfate may be scarcely excreted via urine, and thus accumulate in the blood plasma. Further research is necessary to evaluate the physiological effects of heteroconjugates accumulated in the plasma.

DOI: https://doi.org/10.1039/d1fo03478b

Biomolecules. 2021 Dec 30. pii: 50. [Epub ahead of print]12(1):

Interactions of Pleiotrophin with a Structurally Defined Heparin Hexasaccharide.

Eathen O Ryan, Zhoumai Jiang, Hoa Nguyen, Xu Wang.

Pleiotrophin (PTN) is a potent cytokine that plays an important role in neural generation, angiogenesis, inflammation, and cancers. Its interactions with the polysaccharide glycosaminoglycan (GAG) are crucial to PTN's biological activities. In this study, we investigated the interaction of selectively protonated PTN with the heparin hexasaccharide ΔUA2S-(GlcNS6S-IdoA2S)2-GlcNS6S using solution NMR. The use of a structurally defined oligosaccharide and selectively protonated PTN enabled us to obtain intermolecular contacts using unfiltered NOESY experiments, significantly increasing the amount of high-resolution structural information obtainable. Our data showed that PTN's arginines, lysines, and tryptophans in the two structured domains have strong interactions with the 2-O-sulfated uronate protons in the heparin hexasaccharide. Consistent with the NMR data is the observation that 2-O-desulfation and N-desulfation/N-acetylation significantly decreased heparin hexasaccharides' affinity for PTN, while 6-O-desulfation only modestly affected the interactions with PTN. These results allowed us to hypothesize that PTN has a preference for sulfate clusters centered on the GlcNS6S-IdoA2S disaccharide. Using these data and the fact that PTN domains mostly bind heparin hexasaccharides independently, models of the PTN-heparin complex were constructed.

Keywords: GAG-binding protein; NMR; cytokine; glycosaminoglycan; heparin; pleiotrophin

DOI: https://doi.org/10.3390/biom12010050

Biomolecules. 2022 Jan 05. pii: 77. [Epub ahead of print]12(1):

In-Depth Molecular Dynamics Study of All Possible Chondroitin Sulfate Disaccharides Reveals Key Insight into Structural Heterogeneity and Dynamism.

Balaji Nagarajan, Nehru Viji Sankaranarayanan, Umesh R Desai.

GAGs exhibit a high level of conformational and configurational diversity, which remains untapped in terms of the recognition and modulation of proteins. Although GAGs are suggested to bind to more than 800 biologically important proteins, very few therapeutics have been designed or discovered so far. A key challenge is the inability to identify, understand and predict distinct topologies accessed by GAGs, which may help design novel protein-binding GAG sequences. Recent studies on chondroitin sulfate (CS), a key member of the GAG family, pinpointing its role in multiple biological functions led us to study the conformational dynamism of CS building blocks using molecular dynamics (MD). In the present study, we used the all-atom GLYCAM06 force field for the first time to explore the conformational space of all possible CS building blocks. Each of the 16 disaccharides was solvated in a TIP3P water box with an appropriate number of counter ions followed by equilibration and a production run. We analyzed the MD trajectories for torsional space, inter- and intra-molecular H-bonding, bridging water, conformational spread and energy landscapes. An in-house phi and psi probability density analysis showed that 1→3-linked sequences were more flexible than 1→4-linked sequences. More specifically, phi and psi regions for 1→4-linked sequences were held within a narrower range because of intra-molecular H-bonding between the GalNAc O5 atom and GlcA O3 atom, irrespective of sulfation pattern. In contrast, no such intra-molecular interaction arose for 1→3-linked sequences. Further, the stability of 1→4-linked sequences also arose from inter-molecular interactions involving bridged water molecules. The energy landscape for both classes of CS disaccharides demonstrated increased ruggedness as the level of sulfation increased. The results show that CS building blocks present distinct conformational dynamism that offers the high possibility of unique electrostatic surfaces for protein recognition. The fundamental results presented here will support the development of algorithms that help to design longer CS chains for protein recognition.

Keywords: chondroitin sulfate; conformational analysis; glycosaminoglycans; hydrogen bonding; molecular dynamics; potential energy surface

DOI: https://doi.org/10.3390/biom12010077

Int J Biol Macromol. 2022 Jan 14. pii: S0141-8130(22)00033-2. [Epub ahead of print]202 318-331

Lysine-functionalized chondroitin sulfate improves the biological properties of collagen/chitosan-based injectable hydrogels.

Joanna Klara, Adrianna Marczak, Anna Łatkiewicz, Wojciech Horak, Joanna Lewandowska-Łańcucka.

Novel bioactive collagen/chitosan/lysine-functionalized chondroitin sulfate (CSmod) injectable hydrogels are presented. The modification of CS with amine groups introduced with lysine moieties (the degree of substitution about 21%) guarantees its covalent binding with the hydrogel network while genipin crosslinking. Both the physicochemical and biological features of developed hydrogels might be adjusted by playing with CSmod and crosslinking agent concentrations. It was revealed that materials became more hydrophobic with increased CSmod content, while crosslinking degree and enzymatic degradation studies established the influence of CSmod concentration and Ch:CSmod ratio on the crosslinking process. In situ rheological experiments verified the injectability of resulted systems. The biological in vitro evaluation demonstrated that all designed materials are biocompatible as they supported proliferation and adhesion of MG-63 cell line. In vitro biomineralization study employing simulated body fluid model revealed CSmod-content dependent bioactivity of obtained hydrogels. Importantly for pristine collagen/chitosan materials, the formation of apatite-like structures was not observed. Our findings demonstrate that developed injectable ColChCSmod hydrogels particularly system with the greatest CSmod concentration exhibits high bioactive potential, without the need of applying additional inducers what renders them promising materials within tissue engineering applications.

Keywords: Amine-functionalized chondroitin sulfate; Bioactive injectable hydrogels; Bone tissue engineering

DOI: https://doi.org/10.1016/j.ijbiomac.2022.01.026