bims-supasi 2024-12-08 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2024–12–08
nine papers selected by
Jonathan Wolf Mueller, University of Birmingham

Structural and mechanism-based engineering of sulfotransferase CHST15 for the efficient synthesis of chondroitin sulfate E.
Genetic variability in proteoglycan biosynthetic genes reveals new facets of heparan sulfate diversity.
Sulfation pathways in times of change.
Glycosaminoglycan Adenogenesis Factors: Immunohistochemical Expression in Endometriosis Tissues Compared with the Endometrium.
Inhibition of cell growth and glycosaminoglycan biosynthesis by xylose analog 2-Az-Xyl.
Self-Assembled Nanoparticles with Well-Defined Oligosaccharide Promote Osteogenesis by Regulating Golgi Stress Response.
Loss of carbohydrate sulfotransferase 6 function leads to macular corneal dystrophy phenotypes and skeletal defects in zebrafish.
Steroidogenic pathway in girls diagnosed with autism spectrum disorders.
Analysis of Sulfate Compounds From Sanguisorbae Radix and Its Major Metabolites in Rats by UPLC-MS/MS and Molecular Docking.

Appl Environ Microbiol. 2024 Dec 04. e0157324

Structural and mechanism-based engineering of sulfotransferase CHST15 for the efficient synthesis of chondroitin sulfate E.

Zhonghua Wang, Wei Song, Wanqing Wei, Hejia Qi, Weiwei Meng, Jia Liu, Xiaomin Li, Cong Gao, Liming Liu, Guipeng Hu, Yiwen Zhou, Jing Wu.

  Natural chondroitin sulfate (CS), extracted from animal cartilage, is widely used in the pharmaceuticals and foods. However, contamination with animal-derived heteropolysaccharides presents significant challenges, including potential immune responses. To address this, we developed a green and efficient method for synthesizing chondroitin sulfate E (CSE) via enzymatic synthesis, identifying EcCHST15, a sulfotransferase that catalyzes the conversion of chondroitin sulfate A (CSA) to CSE. We investigated the novel catalytic mechanism of CHST15 through quantum mechanical (QM) calculations and experimental validation, confirming its alignment with the SN2 reaction mechanism. Subsequently, we enhanced the catalytic efficiency of CHST15 using protein engineering, improving the catalytic efficiency from 18.1% in the wild type (WT) to 62.5% in the M7 mutant-a 3.5-fold increase. Finally, we constructed a six-enzyme cascade whole-cell catalyst, achieving a 72.2% conversion of 15 g/L CSA to produce CSE within 24 h. These findings offer a promising strategy for the industrial production of CSE.IMPORTANCECurrent methods for obtaining chondroitin sulfate (CS) primarily rely on tissue extraction and chemical synthesis. However, these approaches are hindered by contamination risks from animal-derived heteropolysaccharides and the technical challenges inherent in complex chemical synthesis, limiting the scalability of industrial CS production. To address this, we developed a green and efficient enzymatic biosynthesis method for chondroitin sulfate E (CSE). By identifying and engineering the sulfotransferase CHST15 from Erpetoichthys calabaricus, we created a mutant (EcCHST15M7) with a 3.5-fold increase in catalytic efficiency toward chondroitin sulfate A (CSA) compared to the wild-type enzyme. Additionally, we constructed a six-enzyme cascade whole-cell biocatalyst, achieving a 72.2% conversion rate from CSA to CSE. This study opens new avenues for the industrial-scale production of CSE through sustainable enzymatic processes.

Keywords:  chondroitin sulfate A; chondroitin sulfate E; enzyme engineering; whole‐cell catalys

DOI:  https://doi.org/10.1128/aem.01573-24
Essays Biochem. 2024 Dec 04. 68(4): 555-578

Genetic variability in proteoglycan biosynthetic genes reveals new facets of heparan sulfate diversity.

Mohand Ouidir Ouidja, Denis S F Biard, Minh Bao Huynh, Xavier Laffray, Wilton Gomez-Henao, Sandrine Chantepie, Gael Le Douaron, Nicolas Rebergue, Auriane Maïza, Heloise Merrick, Aubert De Lichy, Alwyn Dady, Oscar González-Velasco, Karla Rubio, Guillermo Barreto, Kévin Baranger, Valerie Cormier-Daire, Javier De Las Rivas, David G Fernig, Dulce Papy-Garcia.

  Heparan sulfate (HS) and chondroitin sulfate (CS) proteoglycans (PG) consist of a core protein to which the glycosaminoglycan (GAG) chains, HS or CS, are attached through a common linker tetrasaccharide. In the extracellular space, they are involved in the regulation of cell communication, assuring development and homeostasis. The HSPG biosynthetic pathway has documented 51 genes, with many diseases associated to defects in some of them. The phenotypic consequences of this genetic variation in humans, and of genetic ablation in mice, and their expression patterns, led to a phenotypically centered HSPG biosynthetic pathway model. In this model, HS sequences produced by ubiquitous NDST1, HS2ST and HS6ST enzymes are essential for normal development and homeostasis, whereas tissue restricted HS sequences produced by the non-ubiquitous NDST2-4, HS6ST2-3, and HS3ST1-6 enzymes are involved in adaptative behaviors, cognition, tissue responsiveness to stimuli, and vulnerability to disease. The model indicates that the flux through the HSPG/CSPG pathways and its diverse branches is regulated by substrate preferences and protein-protein-interactions. This results in a privileged biosynthesis of HSPG over that of CSPGs, explaining the phenotypes of linkeropathies, disease caused by defects in genes involved in the biosynthesis of the common tetrasaccharide linker. Documented feedback loops whereby cells regulate HS sulfation, and hence the interactions of HS with protein partners, may be similarly implemented, e.g., protein tyrosine sulfation and other posttranslational modifications in enzymes of the HSPG pathway. Together, ubiquitous HS, specialized HS, and their biosynthesis model can facilitate research for a better understanding of HSPG roles in physiology and pathology.

Keywords:  Chondroitin sulfate; Heparan sulfate; biosynthesis; linkeropathies; proteoglycan; sulfation

DOI:  https://doi.org/10.1042/EBC20240106
Essays Biochem. 2024 Dec 04. 68(4): 379-382

Sulfation pathways in times of change.

Jonathan Wolf Mueller, Daniela Fietz, Irundika H K Dias.

  Sulfation pathways are an essential part of overall sulfur metabolism. Sulfation pathways are mainly about sulfate activation, and the making and breaking of biological sulfate esters. This special issue features some extended reflection on what was presented at the SUPA 2023 meeting on Sulfation Pathways. Novel insights into the synthesis and analytics of sulfate, of sulfated conjugates, and of protein persulfides are presented. Oxysterol sulfates are presented as emerging sulfo-metabolites. Sulfation pathways enzymes are discussed in various disease settings. This special issue also presents insights into polysaccharide sulfotransferases and their functional characterization. Finally, cytoplasmic sulfotransferases are highlighted with regards to their impact on DNA-modification, and in the context of endocrine disruptors. All in all, thought-provoking findings, with the potential to guide and stimulate future research in the field of sulfation pathways and beyond.

Keywords:  conjugate analytics; pathways analysis; sulfate activation

DOI:  https://doi.org/10.1042/EBC20230099
Crit Rev Eukaryot Gene Expr. 2025 ;35(1): 67-79

Glycosaminoglycan Adenogenesis Factors: Immunohistochemical Expression in Endometriosis Tissues Compared with the Endometrium.

Pietro Giulio Signorile, Alfonso Baldi, Rosa Viceconte, Emma Carraturo, Mariarosaria Boccellino, Mario Fordellone, Marco Montella.

Endometriosis is a chronic inflammatory pathology estrogen-dependent. It is a condition affecting 5%-10% of women of reproductive age worldwide. Recent evidence indicating an embryological origin of endometriosis has provided new insights into its pathogenesis and potential therapeutic approaches. In this study, we compared the immunohistochemical expression of extracellular matrix molecules involved in the interaction between epithelium and stroma in endometriotic lesions and normal endometrial tissue. A total of 41 cases were analyzed. We examined the immunohistochemical expression of chondroitin sulfate proteoglycan 4 (CSPG4), keratan sulfate, chondroitin sulfate (CS-56), hyaluronic acid, and heparan sulfate (HEP). Our results showed higher expression of CSPG4 and CS-56 in epithelial endometriosis samples compared with normal endometrial tissue, while HEP, keratan sulfate, and hyaluronic acid showed decreased expression in epithelial endometriosis samples relative to normal endometrial tissue. Additionally, endometriotic stroma exhibited more frequent low intensity of hyaluronic acid and HEP compared with normal endometrial stroma. Investigating the levels of these molecules in eutopic and ectopic endometrial tissues enables the identification of potential therapeutic targets, and the development of novel treatments aimed at disrupting the adhesive and invasive properties of endometriotic lesions.

DOI: https://doi.org/10.1615/CritRevEukaryotGeneExpr.2024054273
Biochem Biophys Res Commun. 2024 Nov 27. pii: S0006-291X(24)01619-X. [Epub ahead of print]741 151083

Inhibition of cell growth and glycosaminoglycan biosynthesis by xylose analog 2-Az-Xyl.

Shuji Mizumoto, Kenjiroo Matsumoto, Yuko Tokoro, Naoko Komura, Kazuki Nakajima, Hiromune Ando, Shuhei Yamada, Yasuhiko Kizuka.

  Sugar analogs are versatile chemical tools for probing or inhibiting glycan functions. However, chemical tools are insufficient for glycosaminoglycans (GAGs), which play crucial roles in various biological processes, such as extracellular matrix formation and growth factor signaling. To develop a new compound for detecting GAGs or manipulating GAG functions, we chemically synthesized 2-azide-xylose (2-Az-Xyl), an azide-type analog of Xyl that is a component of the common linkage tetrasaccharide in GAGs, and explored its application to biological experiments. Treatment of cultured cells with 2-Az-Xyl inhibited cell proliferation and reduced the levels of GAGs, particularly heparan sulfate (HS). Although this sugar analog did not perturb the biosynthesis of nucleotide sugars and expression of the key enzymes for HS biosynthesis, 2-Az-Xyl directly inhibited the activity of XYLT2, an initial enzyme for GAG biosynthesis, indicating that 2-Az-Xyl directly inhibits GAG biosynthesis. These findings suggest that 2-Az-Xyl inhibits cell proliferation by blocking GAG biosynthesis through inhibiting XYLT2 activity.

Keywords:  Glycosaminoglycan; Glycosylation; Heparan sulfate; Sugar analog; XYLT2; Xylose

DOI:  https://doi.org/10.1016/j.bbrc.2024.151083
Adv Healthc Mater. 2024 Dec 01. e2402976

Self-Assembled Nanoparticles with Well-Defined Oligosaccharide Promote Osteogenesis by Regulating Golgi Stress Response.

Pingping Niu, Liman Zhao, Jing Yang, Yanan Ding, Xiaoqiao Xu, Sijin Li, Lige Song, Guosong Chen, Yao Sun.

  Osteoporosis, a prevalent disease characterized by low bone density and increased fracture risk, poses significant health challenges for the elderly. Current treatments offer short-term benefits but are limited by long-term efficacy and adverse effects, highlighting the need for new strategies. Chondroitin sulfate polysaccharides (CS), a major component of the bone matrix, are crucial for bone and cartilage health. However, their role in osteoporosis is understudied due to the heterogeneity of natural CS. we found reduced CS levels in osteoporosis patients and developed CS4-NP, a self-assembled tetrasaccharide nanoparticle that mimics CS's structure. CS4-NP, which efficiently delivers the active CS4, significantly improves bone mass in ovariectomized osteoporosis models. It activates the Activating Transcription Factor 4-Cystathionine gamma-Lyase signaling axis in pre-osteoblasts, enhancing osteogenesis. our findings suggest that CS4-NP, an oligosaccharide-based nanomaterial, could address the limitations of current treatments and provide a viable strategy for osteoporosis.

Keywords:  chondroitin sulfate; glyco‐materials; nanoparticles; osteoporosis; self‐assembly

DOI:  https://doi.org/10.1002/adhm.202402976
FEBS J. 2024 Dec 06.

Loss of carbohydrate sulfotransferase 6 function leads to macular corneal dystrophy phenotypes and skeletal defects in zebrafish.

Merve Basol, Esra Ersoz-Gulseven, Helin Ozaktas, Sibel Kalyoncu, Canan Asli Utine, Gulcin Cakan-Akdogan.

  The carbohydrate sulfotransferase 6 (chst6) gene is linked to macular corneal dystrophy (MCD), a rare disease that leads to bilateral blindness due to the accumulation of opaque aggregates in the corneal stroma. chst6 encodes for a keratan sulfate proteoglycan (KSPG) specific sulfotransferase. MCD patients lose sulfated KSPGs (cKS) in the cornea and the serum. The significance of serum cKS loss has not been understood. Zebrafish cornea structure is similar to that of humans and it contains high levels of sulfated cKS in the stroma. Here, zebrafish chst6 is shown to be expressed in the cornea and head structures of the embryos. An animal model of MCD is developed by generating chst6 mutant animals with CRISPR/Cas9-mediated gene editing. The dramatic decrease in cKS epitopes in the mutants was shown with ELISA and immunofluorescence. Morphological defects or alterations of jaw cartilage were detected in a minor fraction of the mutant larvae. Loss of cKS epitopes and morphological defects was fully rescued with wild-type chst6. Mutant adult zebrafish displayed all clinical manifestations of MCD, while a fraction also displayed jaw and skeleton defects. Opaque accumulations formed in the eye, which were alcian blue positive. Loss of cKS in the corneal stroma and a decrease in corneal thickness were shown. Interestingly, alteration of transforming growth factor beta-induced (BIGH3) expression which was not described in patients was also observed. This is the first report of an MCD model in a genetically tractable organism, providing a preclinical model and insight into the importance of KSPG sulfation for proper skeletal morphogenesis.

Keywords:  CRISPR/Cas9; carbohydrate sulfotransferase 6; keratan sulfate proteoglycan; macular corneal dystrophy; zebrafish

DOI:  https://doi.org/10.1111/febs.17337
PLoS One. 2024 ;19(12): e0312933

Steroidogenic pathway in girls diagnosed with autism spectrum disorders.

Katarina Jansakova, Martin Hill, Hana Celusakova, Gabriela Repiska, Marie Bicikova, Ludmila Macova, Katarína Polonyiova, Mária Kopcikova, Daniela Ostatnikova.

The diagnostic prevalence of autism spectrum disorders (ASD) shows boys to be more affected than girls. Due to this reason, there is a lack of research including and observing ASD girls. Present study was aimed to detect hormones of steroidogenesis pathway in prepubertal girls (n = 16) diagnosed with ASD and sex and age matched neurotypical controls (CTRL, n = 16). Collected plasma served for detection of conjugated and unconjugated steroids using gas chromatography tandem-mass spectrometry. We observed higher levels of steroids modulating ionotropic receptors, especially, GABAergic steroids and pregnenolone sulfate in ASD group. Concentration of many steroids throughout the pathway tend to be higher in ASD girls compared to CTRL. Pregnenolone and its isomers together with polar progestins and androstanes, i.e. sulfated steroids, were found to be higher in ASD group in comparison with CTRL group. Based on steroid product to precursor ratios, ASD group showed higher levels of sulfated/conjugated steroids suggesting higher sulfotransferase or lower steroid sulfatase activity and we also obtained data indicating lower activity of steroid 11β-hydroxylase compared to CTRL group despite higher corticosterone level observed in ASD. These findings need to be generalized in future studies to examine both genders and other age groups.

DOI: https://doi.org/10.1371/journal.pone.0312933
Rapid Commun Mass Spectrom. 2025 Feb 28. 39(4): e9947

Analysis of Sulfate Compounds From Sanguisorbae Radix and Its Major Metabolites in Rats by UPLC-MS/MS and Molecular Docking.

Qing Wang, Zhipeng Huo, Bingjing Han, Yujing Sun, Lei Lei, Weitong Xu, Zhaohui Song, Yi He.

   RATIONALE: Sanguisorbae Radix that mainly contains tannins and phenolic compounds has been widely used as a traditional Chinese medicine for treating hemafecia, hemorrhoids metrorrhagia and metrostaxis in clinics. However, there is no report about the sulfate phenolic compounds in Sanguisorbae Radix.
METHODS: Extraction of Sanguisorbae Radix was separated and purified by polyamide resin and octadecyl silane-bonded silica, which were analyzed by HPLC-IT-TOF/MS. Analysis of rat plasma and urine samples using UPLC-Q-TOF/MS. Finally, the newly discovered sulfate esters in Sanguisorbae Radix were investigated by molecular docking to find out the linkage and the possible mechanism to psoriasis.
RESULTS: The chemical structures of methyl-3-O-methylgallate-5-O-sulfate and methyl-3-O-methylgallate were elucidated by the spectral data of HRESIMS and NMR. In addition, 6 sulfate compounds were rapidly identified in Sanguisorbae Radix for the first time, among which 3 sulfate compounds were new natural products. Sixteen prototypes and 11 metabolites were found in rat plasma and urine samples, including 6 sulfate phenolic compounds, and their metabolic pathways in vivo were inferred. Molecular docking technique was used to validate the binding activity of methyl-3-O-methylgallate-5-O-sulfate to psoriasis-related target proteins.
CONCLUSIONS: In this study, sulfate phenolic compounds with high content in Sanguisorbae Radix and biological samples were isolated and characterized by modern spectroscopic techniques, which may provide a theoretical basis for the study of the pharmacological activity and quality control of Sanguisorbae Radix and development of sulfate drugs with a similar structure.

Keywords:  Sanguisorbae Radix; metabolic pathway; molecular docking; sulfate compounds

DOI:  https://doi.org/10.1002/rcm.9947