Synthesis of a chondroitin sulfate disaccharide library and a GAG-binding protein interaction analysis.
01 Apr 2015-Bioorganic & Medicinal Chemistry Letters (Bioorg Med Chem Lett)-Vol. 25, Iss: 7, pp 1407-1411
TL;DR: The common intermediate possessing an orthogonally removable protective group is designed and systematically synthesized all 16 types of CS disaccharide structure generated by sulfation, demonstrating that the 'Sugar Chip' chip technology is effective for the evaluation of binding properties.
About: This article is published in Bioorganic & Medicinal Chemistry Letters.The article was published on 2015-04-01 and is currently open access. It has received 21 citations till now. The article focuses on the topics: Sulfation & Chondroitin sulfate.
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TL;DR: A critical analysis of the tools, molecules, and strategies that can be used to structurally and functionally investigate the formation of chemokine–glycosaminoglycan complexes described to date are provided.
Abstract: Glycosaminoglycans are polysaccharides that occur both at the cell surface and within extracellular matrices. Through their ability to bind to a large array of proteins, almost 500 of which have been identified to date, including most chemokines, these molecules regulate key biologic processes at the cell-tissue interface. To do so, glycosaminoglycans can provide scaffolds to ensure that proteins mediating specific functions will be presented at the correct site and time and can also directly contribute to biologic activities or signaling processes. The binding of chemokines to glycosaminoglycans, which, at the biochemical level, has been mostly studied using heparin, has traditionally been thought of as a mechanism for maintaining haptotactic gradients within tissues along which cells can migrate directionally. Many aspects of chemokine-glycosaminoglycan interactions, however, also suggest that the formation of these complexes could serve additional purposes that go well beyond a simple immobilization process. In addition, progress in glycobiology has revealed that glycosaminoglycan structures, in term of length, sulfation, and epimerization pattern, are specific for cell, tissue, and developmental stage. Glycosaminoglycan regulation and glycosaminoglycan diversity, which cannot be replicated using heparin, thus suggests that these molecules may fine-tune the immune response by selectively recruiting specific chemokines to cell surfaces. In this context, the aim of the present text is to review the chemokine-glycosaminoglycan complexes described to date and provide a critical analysis of the tools, molecules, and strategies that can be used to structurally and functionally investigate the formation of these complexes.
107 citations
Cites methods from "Synthesis of a chondroitin sulfate ..."
...emerging and powerful approach is the use of chemoenzymatic strategies [120], which are based on knowledge of HS bio-...
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TL;DR: The possible impact of alterations in the CS/DS sulfation pattern on the binding capacity and specificity of these GAGs is discussed and potential consequences of the stromal accumulation of chondroitin‐6‐sulfate for the progression and metastasis of cancer are proposed.
Abstract: The remarkable structural heterogeneity of chondroitin sulfate (CS) and dermatan sulfate (DS) generates biological information that can be unique to each of these glycosaminoglycans (GAGs), and changes in their composition are translated into alterations in the binding profiles of these molecules. CS/DS can bind to various cytokines and growth factors, cell surface receptors, adhesion molecules, enzymes and fibrillar glycoproteins of the extracellular matrix, thereby influencing both cell behavior and the biomechanical and biochemical properties of the matrix. In this review, we summarize the current knowledge concerning CS/DS metabolism in the human cancer stroma. The remodeling of the GAG profile in the tumor niche is manifested as a substantial increase in the CS content and a gradual decrease in the proportion between DS and CS. Furthermore, the composition of CS and DS is also affected, which results in a substantial increase in the 6-O-sulfated and/or unsulfated disaccharide content, which is concomitant with a decrease in the 4-O-sulfation level. Here, we discuss the possible impact of alterations in the CS/DS sulfation pattern on the binding capacity and specificity of these GAGs. Moreover, we propose potential consequences of the stromal accumulation of chondroitin-6-sulfate for the progression and metastasis of cancer.
64 citations
TL;DR: An overview of the experimental approaches used in glycosaminoglycomics, of the major GAG-protein interactomes characterized so far, and of the computational tools and databases available to analyze and store GAG structures and interactions is given.
Abstract: Glycosaminoglycans regulate numerous physiopathological processes such as development, angiogenesis, innate immunity, cancer and neurodegenerative diseases. Cell surface GAGs are involved in cell-cell and cell-matrix interactions, cell adhesion and signaling, and host-pathogen interactions. GAGs contribute to the assembly of the extracellular matrix and heparan sulfate chains are able to sequester growth factors in the ECM. Their biological activities are regulated by their interactions with proteins. The structural heterogeneity of GAGs, mostly due to chemical modifications occurring during and after their synthesis, makes the development of analytical techniques for their profiling in cells, tissues, and biological fluids, and of computational tools for mining GAG-protein interaction data very challenging. We give here an overview of the experimental approaches used in glycosaminoglycomics, of the major GAG-protein interactomes characterized so far, and of the computational tools and databases available to analyze and store GAG structures and interactions.
39 citations
Cites background from "Synthesis of a chondroitin sulfate ..."
...CS [75] and HS libraries of oligosaccharides, synthe-...
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TL;DR: CS-D+T, CS-E+A, CS -E+C, and CS- E+E showed greater affinity for basic fibroblast growth factor than did other tetrasaccharides (CS-C+D, C+E, D+D).
25 citations
27 Aug 2020
TL;DR: This review examines aggrecan’s roles in developmental embryonic tissues, in tissues undergoing morphogenetic transition and in mature weight-bearing tissues.
Abstract: This review examines aggrecan’s roles in developmental embryonic tissues, in tissues undergoing morphogenetic transition and in mature weight-bearing tissues. Aggrecan is a remarkably versatile and capable proteoglycan (PG) with diverse tissue context-dependent functional attributes beyond its established role as a weight-bearing PG. The aggrecan core protein provides a template which can be variably decorated with a number of glycosaminoglycan (GAG) side chains including keratan sulphate (KS), human natural killer trisaccharide (HNK-1) and chondroitin sulphate (CS). These convey unique tissue-specific functional properties in water imbibition, space-filling, matrix stabilisation or embryonic cellular regulation. Aggrecan also interacts with morphogens and growth factors directing tissue morphogenesis, remodelling and metaplasia. HNK-1 aggrecan glycoforms direct neural crest cell migration in embryonic development and is neuroprotective in perineuronal nets in the brain. The ability of the aggrecan core protein to assemble CS and KS chains at high density equips cartilage aggrecan with its well-known water-imbibing and weight-bearing properties. The importance of specific arrangements of GAG chains on aggrecan in all its forms is also a primary morphogenetic functional determinant providing aggrecan with unique tissue context dependent regulatory properties. The versatility displayed by aggrecan in biodiverse contexts is a function of its GAG side chains.
22 citations
References
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TL;DR: The findings suggest the possibility of CS-E being a binding partner, a coreceptor, or a genuine receptor for various Hep-binding growth factors in the brain and possibly also in other tissues.
319 citations
TL;DR: All the enzymes in the pathway for synthesis have been cloned, with the exception of the glucuronyl to iduronyL epimerase involved in the formation of dermatan residues.
Abstract: Chondroitin sulfate and dermatan sulfate are synthesized as galactosaminoglycan polymers containing N-acetylgalactosmine alternating with glucuronic acid. The sugar residues are sulfated to varying degrees and positions depending upon the tissue sources and varying conditions of formation. Epimerization of any of the glucuronic acid residues to iduronic acid at the polymer level constitutes the formation of dermatan sulfate. Chondroitin/dermatan glycosaminoglycans are covalently attached by a common tetrasaccharide sequence to the serine residues of core proteins while they are adherent to the inner surface of endoplasmic reticulum/Golgi vesicles. Addition of the first sugar residue, xylose, to core proteins begins in the endoplasmic reticulum, followed by the addition of two galactose residues by two distinct glycosyl transferases in the early cis/medial regions of the Golgi. The linkage tetrasaccharide is completed in the medial/trans Golgi by the addition of the first glucuronic acid residue, followed by transfer of N-acetylgalactosamine to initiate the formation of a galactosaminoglycan rather than a glucosaminoglycan. This specific N-acetylgalactosaminyl transferase is different from the chondroitin synthase involved in generation of the repeating disaccharide units to form the chondroitin polymer. Sulfation of the chondroitin polymer by specific sulfotransferases occurs as the polymer is being formed. All the enzymes in the pathway for synthesis have been cloned, with the exception of the glucuronyl to iduronyl epimerase involved in the formation of dermatan residues.
312 citations
TL;DR: Glycosaminoglycans (GAGs) are complex polysaccharides, which play important roles in cell growth, differentiation, morphogenesis, cell migration, and bacterial/viral infections, and the development of inhibitory drugs for parasitic and viral infections.
Abstract: Glycosaminoglycans (GAGs) are complex polysaccharides, which play important roles in cell growth, differentiation, morphogenesis, cell migration, and bacterial/viral infections. Major GAGs include heparin (Hep)/heparan sulfate, and chondroitin sulfate (CS)/dermatan sulfate (DS). Hep has been used for the treatment of thromboembolic disorders for more than 75 years, and has an established position in therapy today. CS/DS has attracted less attention and its clinical use is limited. However, CS/DS also have intriguing biological activities, which in turn should help in the development of CS/DS-based therapeutics. In this review, the following potential applications of CS/DS chains are discussed. (1) Sugar drugs for parasitic and viral infections. Particular CS variants appear to be involved in infections of various microbes, suggesting that CS/DS oligosaccharide sequences specifically interacting with microbes will lead to the development of inhibitory drugs for these infections. (2) Regenerative medicine. Biological activities of CS/DS chains possibly involve various growth factors, also known as Hep-binding growth factors. Specific CS/DS chains recruit growth/neurotrophic factors and/or potentiate their activities, suggesting that minute amounts of functional CS/DS chains can be utilized for tissue regeneration instead of signaling proteins. (3) Anti-tumor drugs. Specific saccharide structures in CS/DS chains appear to be involved in tumor cell proliferation and metastasis. The detection and identification of such CS/DS saccharide sequences would be an important contribution to cancer therapy.
218 citations
TL;DR: These studies represent the first, direct investigations into the structure-activity relationships of chondroitin sulfate using homogeneous synthetic molecules and define a tetrasaccharide as a minimal motif required for activity.
Abstract: Chondroitin sulfate glycosaminoglycans are sulfated polysaccharides involved in cell division, neuronal development, and spinal cord injury. Here, we report the synthesis and identification of a chondroitin sulfate tetrasaccharide that stimulates the growth and differentiation of neurons. These studies represent the first, direct investigations into the structure−activity relationships of chondroitin sulfate using homogeneous synthetic molecules and define a tetrasaccharide as a minimal motif required for activity.
153 citations
TL;DR: A novel approach for automated solid-phase synthesis of GAG oligosaccharides that is based in part on established methods for generating the glycan portion of glycoproteins and glycolipids is described.
Abstract: Carbohydrates are the most prevalent class of biopolymers on earth. Bound to proteins and lipids, carbohydrates form four structurally and functionally distinct, biologically significant glycoconjugate classes: glycoproteins, glycolipids, glycosylphosphatidylinositol (GPI) anchors, and glycosaminoglycans (GAGs; Figure 1). These structurally diverse macromolecules, which are usually located in the extracellular matrix, are essential for many fundamental cellular processes. GAGs are acidic, negatively charged polysaccharides that transduce extracellular signals to the interior of the cell. Localization is manifested by connection to a transmembrane core protein, to form a proteoglycan (Figure 1). GAGs are highly variable in size, ranging from 20–200 disaccharide repeating units, backbone composition, and the degree and pattern of sulfation. Chondroitin sulfate contains N-acetylb-d-galactosamine and b-d-glucuronic acid and the sulfation and acetylation of particular hydroxy and amino groups varies. The sulfation patterns of GAGs influence the bioactivity of the molecules but limited access to defined GAG structures has impeded mapping structure– activity patterns. Tailor-made GAG oligosaccharides can be synthesized chemically or enzymatically, and they have become valuable for analyzing GAG–protein interactions and their biological relevance. Introducing sulfate groups to specific positions of an oligosaccharide chain adds an additional level of complexity on top of the already challenging synthesis of oligosaccharides. Therefore, currently available methods for the assembly of GAG oligosaccharides, including modular approaches, are time-consuming and lack generality as the synthesis of each target molecule poses an individual challenge. Herein, we describe a novel approach for automated solid-phase synthesis of GAG oligosaccharides that is based in part on established methods for generating the glycan portion of glycoproteins and glycolipids. Key to the success of this procedure was a stable supply of tailor-made differentially protected building blocks, a robust but easily-cleaved linker, to connect the first monosaccharide of the nascent oligosaccharide to the solid support, and the automated synthesizer. A recently-developed automated solid-phase oligosaccharide synthesizer that allows for fully automated, computer-controlled glycan coupling cycles and the introduction of sulfate groups on solid support was further improved to carry out automated sulfation and modification on solid support. Figure 1. Glycoconjugates of the extracellar matrix. Oand N-glycans are linked to proteins by the side chains of serine, threonine, or asparagine. Glycolipids are composed of glycans that are attached to lipids and play an essential role in cellular recognition processes. Glycophosphatidylinositols (GPI) anchor proteins via two fatty acids to the cell membrane. Glycosaminoglycans occur as the glycan side chain in proteoglycans.
142 citations