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Biomolecules Jan 2023Glycosaminoglycans (GAGs) are a class of linear anionic periodic polysaccharides containing disaccharide repetitive units. These molecules interact with a variety of...
Glycosaminoglycans (GAGs) are a class of linear anionic periodic polysaccharides containing disaccharide repetitive units. These molecules interact with a variety of proteins in the extracellular matrix and so participate in biochemically crucial processes such as cell signalling affecting tissue regeneration as well as the onset of cancer, Alzheimer's or Parkinson's diseases. Due to their flexibility, periodicity and chemical heterogeneity, often termed "sulfation code", GAGs are challenging molecules both for experiments and computation. One of the key questions in the GAG research is the specificity of their intermolecular interactions. In this study, we make a step forward to deciphering the "sulfation code" of chondroitin sulfates-4,6 (CS4, CS6, where the numbers correspond to the position of sulfation in NAcGal residue) and dermatan sulfate (DS), which is different from CSs by the presence of IdoA acid instead of GlcA. We rigorously investigate two sets of these GAGs in dimeric, tetrameric and hexameric forms with molecular dynamics-based descriptors. Our data clearly suggest that CS4, CS6 and DS are substantially different in terms of their structural, conformational and dynamic properties, which contributes to the understanding of how these molecules can be different when they bind proteins, which could have practical implications for the GAG-based drug design strategies in the regenerative medicine.
Topics: Dermatan Sulfate; Molecular Dynamics Simulation; Chondroitin Sulfates; Glycosaminoglycans; Sulfates
PubMed: 36830616
DOI: 10.3390/biom13020247 -
Journal of Mass Spectrometry : JMS Mar 2023Biglycan (BGN), a small leucine-rich repeat proteoglycan, is involved in a variety of pathological processes including malignant transformation, for which the...
Biglycan (BGN), a small leucine-rich repeat proteoglycan, is involved in a variety of pathological processes including malignant transformation, for which the upregulation of BGN was found related to cancer cell invasiveness. Because the functions of BGN are mediated by its chondroitin/dermatan sulfate (CS/DS) chains through the sulfates, the determination of CS/DS structure and sulfation pattern is of major importance. In this study, we have implemented an advanced glycomics method based on ion mobility separation (IMS) mass spectrometry (MS) and tandem MS (MS/MS) to characterize the CS disaccharide domains in BGN. The high separation efficiency and sensitivity of this technique allowed the discrimination of five distinct CS disaccharide motifs, of which four irregulated in their sulfation pattern. For the first time, trisulfated unsaturated and bisulfated saturated disaccharides were found in BGN, the latter species documenting the non-reducing end of the chains. The structural investigation by IMS MS/MS disclosed that in one or both of the CS/DS chains, the non-reducing end is 3-O-sulfated GlcA in a rather rare bisulfated motif having the structure 3-O-sulfated GlcA-4-O-sulfated GalNAc. Considering the role played by BGN in cancer cell spreading, the influence on this process of the newly identified sequences will be investigated in the future.
Topics: Chondroitin Sulfates; Biglycan; Tandem Mass Spectrometry; Disaccharides; Dermatan Sulfate; Glycomics
PubMed: 36799777
DOI: 10.1002/jms.4908 -
Regenerative Therapy Mar 2023Synovium-derived mesenchymal stem cells (SMSCs) are multipotential non-hematopoietic progenitor cells that can differentiate into various mesenchymal lineages in adipose...
OBJECTIVE
Synovium-derived mesenchymal stem cells (SMSCs) are multipotential non-hematopoietic progenitor cells that can differentiate into various mesenchymal lineages in adipose and bone tissue, especially in chondrogenesis. Post-transcriptional methylation modifications are relative to the various biological development procedures. N-methyladenosine (mA) methylation has been identified as one of the abundant widespread post-transcriptional modifications. However, the connection between the SMSCs differentiation and mA methylation remains unknown and needs further exploration.
METHODS
SMSCs were derived from synovial tissues of the knee joint of male Sprague-Dawley (SD) rats. In the chondrogenesis of SMSCs, mA regulators were detected by quantitative real-time PCR (RT-PCR) and Western blot (WB). We observed the situation that the knockdown of mA "writer" protein methyltransferase-like (METTL)3 in the chondrogenesis of SMSCs. We also mapped the transcript-wide mA landscape in chondrogenic differentiation of SMSCs and combined RNA-seq and MeRIP-seq in SMSCs by the interference of METTL3.
RESULTS
The expression of mA regulators were regulated in the chondrogenesis of SMSCs, only METTL3 is the most significant factor. In addition, after the knockdown of METTL3, MeRIP-seq and RNA-seq technology were applied to analyze the transcriptome level in SMSCs. 832 DEGs displayed significant changes, consisting of 438 upregulated genes and 394 downregulated genes. DEGs were enriched in signaling pathways regulating the glycosaminoglycan biosynthesis-chondroitin sulfate/dermatan sulfate and ECM-receptor interaction via Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. The findings of this study indicate a difference in transcripts of MMP3, MMP13, and GATA3 containing consensus mA motifs required for methylation by METTL3. Further, the reduction of METTL3 decreased the expression of MMP3, MMP13, and GATA3.
CONCLUSION
These findings confirm the molecular mechanisms of METTL3-mediated mA post-transcriptional change in the modulation of SMSCs differentiating into chondrocytes, thus highlighting the potential therapeutic effect of SMSCs for cartilage regeneration.
PubMed: 36793308
DOI: 10.1016/j.reth.2023.01.005 -
International Journal of Molecular... Jan 2023Chondroitin sulphate (CS) proteoglycans with variable sulphation-motifs along their glycosaminoglycan (GAG) chains are closely associated with the stem cell niche of...
Chondroitin sulphate (CS) proteoglycans with variable sulphation-motifs along their glycosaminoglycan (GAG) chains are closely associated with the stem cell niche of articular cartilage, where they are believed to influence the characteristics of the resident stem cells. Here, we investigated the immunohistochemical distribution of hybrid CS/dermatan sulphate (DS) GAGs in the periphery of the adult chicken cornea, which is the location of the cornea's stem cell niche in a number of species, using a monoclonal antibody, 6C3, that recognises a sulphation motif-specific CS/DS GAG epitope. This revealed positive labelling that was restricted to the subepithelial corneal stroma, as well as nearby bony structures within the sclera, called ossicles. When cultivated on cell culture dishes coated with 6C3-rich CS/DS, corneal stromal cells (keratocytes) that had been isolated from embryonic chicken corneas formed circular colonies, which took several days to reach confluency. A flow cytometric analysis of these keratocytes revealed changes in their expression levels of the indicative stem cell markers, Connexin 43 (Cx43), Paired Box 6 (), B-lymphoma Moloney murine leukemia virus insertion region-1 (Bmi-1), and C-X-C Chemokine Receptor 4 (CXCR4) suggestive of a less-differentiated phenotype compared with expression levels in cells not exposed to CS/DS. These findings support the view that CS/DS promotes the retention of a stem cell phenotype in corneal cells, much as it has been proposed to do in other connective tissues.
Topics: Mice; Chick Embryo; Animals; Chondroitin Sulfates; Proteoglycans; Glycosaminoglycans; Stem Cells; Cornea
PubMed: 36768414
DOI: 10.3390/ijms24032095 -
Clinica Chimica Acta; International... Feb 2023Mucopolysaccharidoses (MPSs) are inherited genetic diseases caused by an absence or deficiency of lysosomal enzymes responsible for catabolizing glycosaminoglycans...
BACKGROUND
Mucopolysaccharidoses (MPSs) are inherited genetic diseases caused by an absence or deficiency of lysosomal enzymes responsible for catabolizing glycosaminoglycans (GAGs). Undiagnosed patients, or those without adequate treatment in early life, can be severely and irreversibly affected by the disease. In this study, we applied liquid chromatography-high resolution mass spectrometry (LC-HRMS)-based untargeted metabolomics to identify potential biomarkers for MPS disorders to better understand how MPS may affect the metabolome of patients.
METHODS
Urine samples from 37 MPS patients (types I, II, III, IV, and VI; untreated and treated with enzyme replacement therapy (ERT)) and 38 controls were analyzed by LC-HRMS. Data were processed by an untargeted metabolomics workflow and submitted to multivariate statistical analyses to reveal significant differences between the MPS and control groups.
RESULTS
A total of 12 increased metabolites common to all MPS types were identified. Dipeptides, amino acids and derivatives were increased in the MPS group compared to controls. N-acetylgalactosamines 4- or 6-sulfate, important constituents of GAGs, were also elevated in MPS patients, most prominently in those with MPS VI. Notably, treated patients exhibited lower levels of the aforementioned acylaminosugars than untreated patients in all MPS types.
CONCLUSIONS
Untargeted metabolomics has enabled the detection of metabolites that could improve our understanding of MPS physiopathology. These potential biomarkers can be utilized in screening methods to support diagnosis and ERT monitoring.
Topics: Humans; Heparitin Sulfate; Dermatan Sulfate; Tandem Mass Spectrometry; Mucopolysaccharidoses; Glycosaminoglycans; Chromatography, Liquid; Metabolomics; Biomarkers
PubMed: 36764508
DOI: 10.1016/j.cca.2023.117250 -
Glycoconjugate Journal Apr 2023Dried leech (Whitmania pigra whitman) has been widely used as a traditional animal-based Chinese medicine. Dried leech extracts have been reported to have various...
Dried leech (Whitmania pigra whitman) has been widely used as a traditional animal-based Chinese medicine. Dried leech extracts have been reported to have various biological activities that are often associated with mammalian glycosaminoglycans. However, their presence and possible structural characteristics within dried leech were previously unknown. In this study, glycosaminoglycans were isolated from dried leech for the first time and their structures were analyzed by the combination of Fourier-transform infrared spectroscopy, liquid chromatography-ion trap/time-of-flight mass spectrometry and polyacrylamide gel electrophoresis. Heparan sulfate and chondroitin sulfate/dermatan sulfate were detected in dried leech with varied disaccharide compositions and possess a heterogeneous structure. Heparan sulfate species possess an equal amount of total 2-O-sulfated, N-sulfated and acetylated disaccharides, while chondroitin sulfate /dermatan sulfate contain high content of 4-O-sulfated disaccharides. Also, the quantitative analysis revealed that the contents of heparan sulfate and chondroitin/dermatan sulfate in dried leech varied significantly, with chondroitin/dermatan sulfate being by far the most abundant. This novel structural information could help clarify the possible involvement of these polysaccharides in the biological activities of the dried leech. Furthermore, leech glycosaminoglycans showed a strong ABTS radical scavenging ability, which suggests the potential of leech polysaccharides for exploitation in the nutraceutical and pharmaceutical industries.
Topics: Animals; Glycosaminoglycans; Chondroitin Sulfates; Dermatan Sulfate; Antioxidants; Heparitin Sulfate; Mammals; Disaccharides
PubMed: 36749437
DOI: 10.1007/s10719-023-10105-y -
Cureus Jan 2023Hurler syndrome (HS) belongs to the category of mucopolysaccharidosis (MPS), a spectrum of rare genetic disorders of the mucopolysaccharides metabolism. This syndrome is...
Hurler syndrome (HS) belongs to the category of mucopolysaccharidosis (MPS), a spectrum of rare genetic disorders of the mucopolysaccharides metabolism. This syndrome is due to a defect in α-iduronidase, an enzyme responsible for the degradation of the glycosaminoglycans (GAGs) heparin and dermatan sulfate. Intra and extracellular accumulation of these non-metabolized substances may lead to multisystemic dysfunction, with severe stomatognathic involvement that may often need treatment. The aim of this article is to present the heterogeneity of orofacial and radiographic findings observed in two patients with HS with long-term follow-up, who were referred to our Stomatology department.
PubMed: 36741627
DOI: 10.7759/cureus.33313 -
Carbohydrate Polymers Apr 2023The sulfation patterns of chondroitin sulfate (CS)/dermatan sulfate (DS), which encode unique biological information, play critical roles in the various biological...
The sulfation patterns of chondroitin sulfate (CS)/dermatan sulfate (DS), which encode unique biological information, play critical roles in the various biological functions of CS/DS chains. CS/DS sulfatases, which can specifically hydrolyze sulfate groups, could potentially be essential tools for deciphering and changing the biological information encoded by these sulfation patterns. However, endosulfatase with high activity to efficiently hydrolyze the sulfate groups inside CS/DS polysaccharides have rarely been identified, which hinders the practical applications of CS/DS sulfatases. Herein, a novel CS/DS 4-O-endosulfatase (endoBI4SF) with a strong ability to completely remove 4-O-sulfated groups inside various CS/DS polysaccharides was identified and successfully used to investigate the biological roles of 4-O-sulfated CS/DS in vitro and in vivo. This study provides a much-needed tool to tailor the sulfation patterns and explore the related functions of 4-O-sulfated CS/DS chains in vitro and in vivo.
Topics: Chondroitin Sulfates; Dermatan Sulfate; Polysaccharides; Sulfatases
PubMed: 36737182
DOI: 10.1016/j.carbpol.2022.120508