The Enzymatic Degradation of Heparan Sulfate

Griffin, Laura Susan; Gloster, T.M.

doi:10.2174/0929866524666170724113452

Cited by 16 publications

(17 citation statements)

References 145 publications

(169 reference statements)

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“…from 6S-GlcNAc) of HS. On the other hand, the turnover of GAGs in cells is achieved by exo-acting lysosomal enzymes, a series of specific enzymes including glycosidases and sulfatases, whose sequential action degrades the target GAG into monosaccharide units and sulfate anions [15]. Importantly, impairment of these enzymes leads to the accumulation of partial breakdown products, resulting in a series of metabolic disorders termed mucopolysaccharidoses (MPS) [16,17].…”

Section: Introductionmentioning

confidence: 99%

Mobility shift-based electrophoresis coupled with fluorescent detection enables real-time enzyme analysis of carbohydrate sulfatase activity

Byrne

London

Eyers

et al. 2021

Biochemical Journal

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Sulfated carbohydrate metabolism is a fundamental process, which occurs in all domains of life. Carbohydrate sulfatases are enzymes that remove sulfate groups from carbohydrates and are essential to the depolymerisation of complex polysaccharides. Despite their biological importance, carbohydrate sulfatases are poorly studied and challenges remain in accurately assessing the enzymatic activity, specificity and kinetic parameters. Most notably, separation of desulfated products from sulfated substrates is currently a time-consuming process. In this paper, we describe the development of rapid capillary electrophoresis coupled to substrate fluorescence detection as a high-throughput and facile means of analysing carbohydrate sulfatase activity. The approach has utility for the determination of both kinetic and inhibition parameters and is based on existing microfluidic technology coupled to a new synthetic fluorescent 6S-GlcNAc carbohydrate substrate. Furthermore, we compare this technique, in terms of both time and resources, to high performance anion exchange chromatography and NMR-based methods, which are the two current ‘gold standards’ for enzymatic carbohydrate sulfation analysis. Our study clearly demonstrates the advantages of mobility shift assays for the quantification of near real-time carbohydrate desulfation by purified sulfatases, and will support the search for small molecule inhibitors of these disease-associated enzymes.

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Section: Introductionmentioning

confidence: 99%

Mobility shift-based electrophoresis coupled with fluorescent detection enables real-time enzyme analysis of carbohydrate sulfatase activity

Byrne

London

Eyers

et al. 2021

Biochemical Journal

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“…Different glioblastoma primary cell cultures have been shown to possess a high heterogeneity of the HS structure and composition, which has been associated with the adhesive properties and invasive activity of these cells [ 15 ]. However, molecular mechanisms of deregulation of HS in gliomas remain poorly understood, and appear to be determined by the balance of the processes of biosynthesis of HS and its extracellular degradation [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

Heparan Sulfate Biosynthetic System Is Inhibited in Human Glioma Due to EXT1/2 and HS6ST1/2 Down-Regulation

Ushakov

Tsidulko

Bourdonnaye

et al. 2017

IJMS

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Heparan sulfate (HS) is an important component of the extracellular matrix and cell surface, which plays a key role in cell–cell and cell–matrix interactions. Functional activity of HS directly depends on its structure, which determined by a complex system of HS biosynthetic enzymes. During malignant transformation, the system can undergo significant changes, but for glioma, HS biosynthesis has not been studied in detail. In this study, we performed a comparative analysis of the HS biosynthetic system in human gliomas of different grades. RT-PCR analysis showed that the overall transcriptional activity of the main HS biosynthesis-involved genes (EXT1, EXT2, NDST1, NDST2, GLCE, HS2ST1, HS3ST1, HS3ST2, HS6ST1, HS6ST2, SULF1, SULF2, HPSE) was decreased by 1.5–2-fold in Grade II-III glioma (p < 0.01) and by 3-fold in Grade IV glioma (glioblastoma multiforme, GBM) (p < 0.05), as compared with the para-tumourous tissue. The inhibition was mainly due to the elongation (a decrease in EXT1/2 expression by 3–4-fold) and 6-O-sulfation steps (a decrease in 6OST1/2 expression by 2–5-fold) of the HS biosynthesis. Heparanase (HPSE) expression was identified in 50% of GBM tumours by immunostaining, and was characterised by a high intratumoural heterogeneity of the presence of the HPSE protein. The detected disorganisation of the HS biosynthetic system in gliomas might be a potential molecular mechanism for the changes of HS structure and content in tumour microenvironments, contributing to the invasion of glioma cells and the development of the disease.

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“…The elongation process starts with the addition of one N-acetyl glucosamine (GlcNAc) to the linkage region, step under control of the EXTL2 and EXTL3 genes, whose products (EXTL2 and EXTL3) have GlcNAc-transferase I activity [ 24 ]. After this initiation step with the participation of the EXTL genes, the elongation of the HS chain takes place by the action of the EXT1–EXT2 complex, which alternatively adds glucuronic acid (GlcA) and GlcNAc residues to the chain, forming polymers of different length [ 26 ]. Mutations in EXT1 and EXT2 result in reduced HS in the cartilage and cause hereditary multiple exostoses, an autosomal dominant disorder affecting skeleton with a risk of malignant transformation [ 27 ].…”

Section: Heparan Sulfatementioning

confidence: 99%

“…HS is degraded within the lysosomes ( Figure 1 ), to which it arrives through the endosomal pathway [ 26 ]. First, in the extracellular matrix, some endosulfatases and a secreted heparanase could partially degrade HS chains, giving rise to smaller fragments.…”

Section: Heparan Sulfatementioning

confidence: 99%

“…After the fragmentation, the following enzymes proceed with the complete degradation of the small fragments by acting sequentially ( Figure 1 ): iduronate 2-sulfatase (IDS), α-L-iduronidase (IDUA), heparan N-sulfatase or sulfamidase (SGSH, mutated in Sanfilippo syndrome type A), acetyl-CoA α-glucosaminide N-acetyltransferase (HGSNAT, mutated in Sanfilippo syndrome type C), α-N-acetylglucosaminidase (NAGLU, mutated in Sanfilippo syndrome type B), glucuronate 2-sulfatase (GDS), β-glucuronidase (GUSB), and N-acetylglucosamine 6-sulfatase (GNS, mutated in Sanfilippo syndrome type D) [ 26 ]. It has been suggested that all these enzymes function as a complex in the lysosomes [ 28 ].…”

Section: Heparan Sulfatementioning

confidence: 99%

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Sanfilippo Syndrome: Molecular Basis, Disease Models and Therapeutic Approaches

Benetó

Vilageliu

Grinberg

et al. 2020

IJMS

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Sanfilippo syndrome or mucopolysaccharidosis III is a lysosomal storage disorder caused by mutations in genes responsible for the degradation of heparan sulfate, a glycosaminoglycan located in the extracellular membrane. Undegraded heparan sulfate molecules accumulate within lysosomes leading to cellular dysfunction and pathology in several organs, with severe central nervous system degeneration as the main phenotypical feature. The exact molecular and cellular mechanisms by which impaired degradation and storage lead to cellular dysfunction and neuronal degeneration are still not fully understood. Here, we compile the knowledge on this issue and review all available animal and cellular models that can be used to contribute to increase our understanding of Sanfilippo syndrome disease mechanisms. Moreover, we provide an update in advances regarding the different and most successful therapeutic approaches that are currently under study to treat Sanfilippo syndrome patients and discuss the potential of new tools such as induced pluripotent stem cells to be used for disease modeling and therapy development.

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The Enzymatic Degradation of Heparan Sulfate

Cited by 16 publications

References 145 publications

Mobility shift-based electrophoresis coupled with fluorescent detection enables real-time enzyme analysis of carbohydrate sulfatase activity

Mobility shift-based electrophoresis coupled with fluorescent detection enables real-time enzyme analysis of carbohydrate sulfatase activity

Heparan Sulfate Biosynthetic System Is Inhibited in Human Glioma Due to EXT1/2 and HS6ST1/2 Down-Regulation

Sanfilippo Syndrome: Molecular Basis, Disease Models and Therapeutic Approaches

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