A Chondroitin Sulfate Small Molecule that Stimulates Neuronal Growth

Tully, Sarah E.; Mabon, Ross; Gama, Cristal I.; Tsai, Sherry M.; Liu, Xuewei; Hsieh‐Wilson, Linda C.

doi:10.1021/ja0484045

Cited by 149 publications

(136 citation statements)

References 19 publications

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“…2C). However, the ability of BDNF and other NTs to bind weakly to other sulfation motifs was unexpected, given that only the CS-E motif stimulated neurite outgrowth (22,39). Thus, we postulated that CS-E might interact with additional proteins, possibly forming proteinprotein complexes between NTs and their receptors and that the formation of such complexes might impart greater selectivity for the CS-E motif.…”

Section: Identification Of New Glycosaminoglycan-protein Interactionsmentioning

confidence: 94%

“…The NTs have critical functions in many aspects of neuronal development, including neurite outgrowth, cell survival, differentiation, and proliferation (36,37 the adult nervous system (36-38) and have been implicated in neurodegenerative diseases (36). Previously, we found that a tetrasaccharide containing a specific sulfation motif, CS-E, stimulates the outgrowth of developing hippocampal neurons (22,39). Our studies implicated brain-derived neurotrophic factor (BDNF) as one of the proteins responsible for mediating the effects of CS-E (22).…”

Section: Identification Of New Glycosaminoglycan-protein Interactionsmentioning

confidence: 99%

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Elucidating glycosaminoglycan–protein–protein interactions using carbohydrate microarray and computational approaches

Rogers

Clark

Tully

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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100

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Glycosaminoglycan polysaccharides play critical roles in many cellular processes, ranging from viral invasion and angiogenesis to spinal cord injury. Their diverse biological activities are derived from an ability to regulate a remarkable number of proteins. However, few methods exist for the rapid identification of glycosaminoglycan-protein interactions and for studying the potential of glycosaminoglycans to assemble multimeric protein complexes. Here, we report a multidisciplinary approach that combines new carbohydrate microarray and computational modeling methodologies to elucidate glycosaminoglycan-protein interactions. The approach was validated through the study of known protein partners for heparan and chondroitin sulfate, including fibroblast growth factor 2 (FGF2) and its receptor FGFR1, the malarial protein VAR2CSA, and tumor necrosis factor-α (TNF-α). We also applied the approach to identify previously undescribed interactions between a specific sulfated epitope on chondroitin sulfate, CS-E, and the neurotrophins, a critical family of growth factors involved in the development, maintenance, and survival of the vertebrate nervous system. Our studies show for the first time that CS is capable of assembling multimeric signaling complexes and modulating neurotrophin signaling pathways. In addition, we identify a contiguous CS-E-binding site by computational modeling that suggests a potential mechanism to explain how CS may promote neurotrophin-tyrosine receptor kinase (Trk) complex formation and neurotrophin signaling. Together, our combined microarray and computational modeling methodologies provide a general, facile means to identify new glycosaminoglycan-protein-protein interactions, as well as a molecular-level understanding of those complexes. G lycosaminoglycans (GAGs) regulate a wide range of physiological processes, including viral invasion, blood coagulation, cell growth, and spinal cord injury (1-4). Assembled from repeating disaccharide units, GAGs display diverse patterns of sulfation (SI Appendix, Fig. S1). These sulfation patterns are believed to have important functional consequences, enabling the polysaccharides to interact with a wide variety of proteins (1, 2). However, the precise sulfation motifs involved in protein recognition are understood in only a few cases (1,4,5). Moreover, studies of heparan sulfate (HS) interactions with fibroblast growth factors suggest that GAGs can assist in the assembly of multimeric protein complexes, thereby modulating signal transduction pathways (6-10). Yet, only a few such examples have been elucidated, and the extent to which other GAGs such as chondroitin sulfate (CS) engage in the formation of multimeric protein complexes remains unknown. Elucidating the interactions of specific GAG substructures with proteins and large protein-protein complexes will be critical for understanding the structure-activity relationships of GAGs and the mechanisms underlying important biological processes.Several methods have been developed to study GAG-protein interac...

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Section: Identification Of New Glycosaminoglycan-protein Interactionsmentioning

confidence: 94%

Section: Identification Of New Glycosaminoglycan-protein Interactionsmentioning

confidence: 99%

Elucidating glycosaminoglycan–protein–protein interactions using carbohydrate microarray and computational approaches

Rogers

Clark

Tully

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

100

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“…3,4 In this solution-based assay, a monovalent tetrasaccharide at 0.5 µg/mL glucuronic acid concentration had minimal activity (Supporting Information). In contrast, glycopolymer 16 at the same glucuronic acid concentration exhibited maximal activity, inducing neurite inhibition to the same extent as the natural polysaccharide ( Figure 2).…”

Section: Scheme 3 Cs Glycomimetics Amentioning

confidence: 95%

Neuroactive Chondroitin Sulfate Glycomimetics

et al. 2008

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Chondroitin sulfate (CS) glycosaminoglycans play critical roles in various physiological processes, including cell division, inflammation, and spinal cord injury. 1 However, the complexity of this important class of molecules has hindered efforts to relate structure to function and to generate defined molecular tools for manipulating CS activity. Comprising 40-200 sulfated disaccharide units, CS is thought to contain "blocks" of high and low sulfation, with highly sulfated regions serving as binding sites for growth factors, cytokines, and other proteins. 2,3 Although a tetrasaccharide is often sufficient for molecular recognition and neuronal activity, 3,4 longer sequences may further enhance protein binding and allow for the assembly of multimeric protein complexes. 5 Given the challenges inherent in the synthesis of large oligosaccharides, we sought the development of CS mimetics that would retain key properties of CS polysaccharides, yet be more synthetically accessible. Here, we describe the first synthesis and characterization of CS glycopolymers with defined sulfation sequence and tunable chemical and biological properties.Surprisingly few polymers based on glycosaminoglycan structures have been reported. Heparin-like glycopolymers have been synthesized from simple monosaccharides such as N-acetyl-Dglucosamine. 6 However, no high molecular weight glycopolymers have been assembled from disaccharide building blocks found in heparin/heparan sulfate, chondroitin, dermatan, or keratan sulfate glycosaminoglycans. 7 We envisioned that such glycopolymers would more closely mimic the natural polysaccharides (Figure 1), facilitating explorations into the importance of macromolecular structure (e.g., distance, number, and orientation between proteinbinding epitopes, multivalency) and providing tools to perturb the functions of CS in vivo.The functional complexity of CS represents a major challenge to the synthesis of glycopolymers as hydroxyl, carboxylic acid, and sulfate groups are reactive under cationic and anionic polymerizations. As such, we explored ruthenium-catalyzed ring-opening metathesis polymerization (ROMP), 8 which has been shown to tolerate a broad range of functional groups and affords glycopolymers of controllable length and narrow polydispersity. 9 In place of norbornene derivatives used previously, 9 we chose cis-cyclooctene monomers, as less constrained scaffolds have been reported to adapt better to biological receptors. 10 To ensure a high degree of control over the sulfation pattern, sulfated monomers were used in the polymerization reaction.We first probed the efficiency of ROMP using disaccharides 6 and 7, which display the biologically active CS-E sulfation motif (Scheme 1). To obtain 6 and 7, trichloroacetimidate 11 donor 1 4 was coupled to the cyclooctene acceptor 2 to afford exclusively the -linked disaccharide 3 in 69% yield. Removal of the tertbutyldimethylsilyl group using HF‚pyridine gave 4, which served as a glycosyl acceptor to assemble tetrasaccharides (Scheme 2). Free radical...

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“…Further studies are needed to investigate the GlcUA/IdoUA distribution pattern. The purified CS-H preparation contained substantial proportions of both E and iE units, the importance of which has been suggested in the growth factor-binding (13,45) and neuritogenic (11,14,46) activities of CS and DS and which needs to be proven in such activities of CS-H. The iE unit has been identified in DS from mammalian liver (47), bovine kidney (48), and embryonic and adult pig brains (16).…”

Section: Table III Disaccharide Composition Of Purified Cs-hmentioning

confidence: 99%

Structural and Functional Characterization of Oversulfated Chondroitin Sulfate/Dermatan Sulfate Hybrid Chains from the Notochord of Hagfish

Nandini

Mikami

Ohta

et al. 2004

Journal of Biological Chemistry

115

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Pivotal functions of glycosaminoglycan (GAG) 1 side chains of proteoglycans (PGs), especially heparan sulfate (HS), have been implicated in biological processes such as cell adhesion, proliferation, and differentiation and tissue morphogenesis (1). Chondroitin sulfate (CS) and dermatan sulfate (DS) are also found ubiquitously in the extracellular matrices and at cell surfaces, being main components of cartilage and skin, respectively. CS consists of repeating disaccharide units of -4Gl-cUA␤1-3GalNAc␤1-, whereas DS is an isomeric form of CS and is formed from precursor CS through the action of glucuronyl C5 epimerase, thus consisting of disaccharide units of -4Gl-cUA␤1-3GalNAc␤1-and -4IdoUA␣1-3GalNAc␤1-in varying proportions (2). These disaccharide units are modified during chain elongation by specific sulfotransferases at C-2 of GlcUA/ IdoUA and/or C-4 and/or C-6 of GalNAc in various combinations, producing characteristic sulfation patterns critical for binding to various functional proteins displaying enormous structural diversity, comparable with that of HS, by embedding multiple overlapping functional sequences (3). Sulfation profiles of GAGs change during development (4, 5). Growing evidence indicates the involvement of CS and DS in the signaling of various heparin-binding growth factors and cytokines (6 -8).We and others have shown the importance of this class of molecule, from simple chondroitin involved in cell division of a nematode (9) to differentially oversulfated CS-D and CS-E involved in neuroregulatory functions (10 -12) and the binding of growth factors in mammalian systems (13). While pursuing the critical structural elements in the CS variants, we became *

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A Chondroitin Sulfate Small Molecule that Stimulates Neuronal Growth

Cited by 149 publications

References 19 publications

Elucidating glycosaminoglycan–protein–protein interactions using carbohydrate microarray and computational approaches

Elucidating glycosaminoglycan–protein–protein interactions using carbohydrate microarray and computational approaches

Neuroactive Chondroitin Sulfate Glycomimetics

Structural and Functional Characterization of Oversulfated Chondroitin Sulfate/Dermatan Sulfate Hybrid Chains from the Notochord of Hagfish

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