The Experts below are selected from a list of 5115 Experts worldwide ranked by ideXlab platform
Kazuyuki Sugahara - One of the best experts on this subject based on the ideXlab platform.
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molecular interactions between chondroitin Dermatan Sulfate and growth factors receptors matrix proteins
Current Opinion in Structural Biology, 2015Co-Authors: Shuji Mizumoto, Kazuyuki Sugahara, Shuhei YamadaAbstract:Recent functional studies on chondroitin Sulfate-Dermatan Sulfate (CS-DS) demonstrated its indispensable roles in various biological events including brain development and cancer. CS-DS proteoglycans exert their physiological activity through interactions with specific proteins including growth factors, cell surface receptors, and matrix proteins. The characterization of these interactions is essential for regulating the biological functions of CS-DS proteoglycans. Although amino acid sequences on the bioactive proteins required for these interactions have already been elucidated, the specific saccharide sequences involved in the binding of CS-DS to target proteins have not yet been sufficiently identified. In this review, recent findings are described on the interaction between CS-DS and some proteins which are especially involved in the central nervous system and cancer development/metastasis.
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Dermatan Sulfate in tunicate phylogeny order specific sulfation pattern and the effect of 4idoa 2 Sulfate β 1 3galnac 4 Sulfate β 1 motifs in Dermatan Sulfate on heparin cofactor ii activity
BMC Biochemistry, 2011Co-Authors: Eliene O. Kozlowski, P C Lima, Tito Monteiro Da Cruz Lotufo, Cristina P Vicente, Kazuyuki Sugahara, Mauro S G PavaoAbstract:Previously, we have reported the presence of highly Sulfated Dermatans in solitary ascidians from the orders Phlebobranchia (Phallusia nigra) and Stolidobranchia (Halocynthia pyriformis and Styela plicata). Despite the identical disaccharide backbone, consisting of [→4IdoA(2S)β-1→3GalNAcβ-1→], those polymers differ in the position of sulfation on the N-Acetyl galactosamine, which can occur at carbon 4 or 6. We have shown that position rather than degree of sulfation is important for heparin cofactor II activity. As a consequence, 2,4- and 2,6-Sulfated Dermatans have high and low heparin cofactor II activities, respectively. In the present study we extended the disaccharide analysis of ascidian Dermatan Sulfates to additional species of the orders Stolidobranchia (Herdmania pallida, Halocynthia roretzi) and Phlebobranchia (Ciona intestinalis), aiming to investigate how sulfation evolved within Tunicata. In addition, we analysed how heparin cofactor II activity responds to Dermatan Sulfates containing different proportions of 2,6- or 2,4-diSulfated units. Disaccharide analyses indicated a high content of diSulfated disaccharide units in the Dermatan Sulfates from both orders. However, the degree of sulfation decreased from Stolidobranchia to Phlebobranchia. While 76% of the disaccharide units in Dermatan Sulfates from stolidobranch ascidians are diSulfated, 53% of diSulfated disaccharides are found in Dermatan Sulfates from phlebobranch ascidians. Besides this notable difference in the sulfation degree, Dermatan Sulfates from phlebobranch ascidians contain mainly 2,6-Sulfated disaccharides whereas Dermatan Sulfate from the stolidobranch ascidians contain mostly 2,4-Sulfated disaccharides, suggesting that the biosynthesis of Dermatan Sulfates might be differently regulated during tunicates evolution. Changes in the position of sulfation on N-acetylgalactosamine in the disaccharide [→4IdoA(2-Sulfate)β-1→3GalNAcβ-1→] modulate heparin cofactor II activity of Dermatan Sulfate polymers. Thus, high and low heparin cofactor II stimulating activity is observed in 2,4-Sulfated Dermatan Sulfates and 2,6-Sulfated Dermatan Sulfates, respectively, confirming the clear correlation between the anticoagulant activities of Dermatan Sulfates and the presence of 2,4-Sulfated units. Our results indicate that in ascidian Dermatan Sulfates the position of sulfation on the GalNAc in the disaccharide [→4IdoA(2S)β-1→3GalNAcβ-1→] is directly related to the taxon and that the 6-O sulfation is a novelty apparently restricted to the Phlebobranchia. We also show that the increased content of [→4IdoA(2S)β-1→3GalNAc(4S)β-1→] disaccharide units in Dermatan Sulfates from Stolidobranchia accounts for the increased heparin cofactor II stimulating activity.
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potential therapeutic application of chondroitin Sulfate Dermatan Sulfate
Current Drug Discovery Technologies, 2008Co-Authors: Shuhei Yamada, Kazuyuki SugaharaAbstract: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.
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overSulfated Dermatan Sulfate exhibits neurite outgrowth promoting activity toward embryonic mouse hippocampal neurons implications of Dermatan Sulfate in neuritogenesis in the brain
Journal of Biological Chemistry, 2003Co-Authors: Megumi Hikino, Mauro S G Pavao, Tadahisa Mikami, Andreas Faissner, Anacristina E S Vilelasilva, Kazuyuki SugaharaAbstract:Brain-specific chondroitin Sulfate (CS) proteoglycan (PG) DSD-1-PG/6B4-PG/phosphacan isolated from neonatal mouse brains exhibits neurite outgrowth-promoting activity toward embryonic rat and mouse hippocampal neurons in vitro through the so-called DSD-1 epitope embedded in its glycosaminoglycan side chains. OverSulfated CS variants, CS-D from shark cartilage and CS-E from squid cartilage, also possess similar activities. We have proposed that the neuritogenic property of the DSD-1 epitope may be attributable to a distinct CS structure characterized by the diSulfated D disaccharide unit [GlcUA(2S)-GalNAc(6S)]. In this study, we assessed neuritogenic potencies of various overSulfated Dermatan Sulfate (DS) preparations purified from hagfish notochord, the bodies of two kinds of ascidians and embryonic sea urchin, which are characterized by the predominant diSulfated disaccharide units of [IdoUA-GalNAc(4S,6S)] (68%), [IdoUA(2S)-GalNAc(4S)] (66%) plus [IdoUA(2S)-GalNAc(6S)] (5%), [IdoUA(2S)-GalNAc (6S)] (>90%), and [IdoUA-GalNAc(4S,6S)] (74%), respectively. They exerted marked neurite outgrowth-promoting activities, resulting in distinct morphological features depending on the individual structural features. Such activities were not observed for a less Sulfated DS preparation derived from porcine skin, which has a monoSulfated disaccharide unit [IdoUA-Gal-NAc(4S)] as a predominant unit. The neurite outgrowth-promoting activities of these overSulfated DS preparations and DSD-1-PG were eliminated by the specific enzymatic cleavage of GalNAc-IdoUA linkages characteristic of DS using chondroitinase B. In addition, chemical analysis of the glycosaminoglycan side chains of DSD-1-PG revealed the DS-type structures. These observations suggest potential novel neurobiological functions of overSulfated DS structures and may reflect the physiological neuritogenesis during brain development by mammalian overSulfated DS structures exemplified by the DSD-1 epitope.
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recent advances in the structural biology of chondroitin Sulfate and Dermatan Sulfate
Current Opinion in Structural Biology, 2003Co-Authors: Kazuyuki Sugahara, Toru Uyama, Tadahisa Mikami, Souhei Mizuguchi, Kazuya Nomura, Hiroshi KitagawaAbstract:Recent glycobiology studies have suggested fundamental biological functions for chondroitin, chondroitin Sulfate and Dermatan Sulfate, which are widely distributed as glycosaminoglycan sidechains of proteoglycans in the extracellular matrix and at cell surfaces. They have been implicated in the signaling functions of various heparin-binding growth factors and chemokines, and play critical roles in the development of the central nervous system. They also function as receptors for various pathogens. These functions are closely associated with the sulfation patterns of the glycosaminoglycan chains. Surprisingly, nonSulfated chondroitin is indispensable in the morphogenesis and cell division of Caenorhabditis elegans, as revealed by RNA interference experiments of the recently cloned chondroitin synthase gene and by the analysis of mutants of squashed vulva genes.
Richard L Gallo - One of the best experts on this subject based on the ideXlab platform.
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exogenous addition of a c xylopyranoside derivative stimulates keratinocyte Dermatan Sulfate synthesis and promotes migration
PLOS ONE, 2011Co-Authors: Jun Muto, Nandita Natasha Naidu, Kenshi Yamasaki, Nathalie Pineau, Lionel Breton, Richard L GalloAbstract:As C-Xyloside has been suggested to be an initiator of glycosaminoglycan (GAG) synthesis, and GAGs such as Dermatan Sulfate (DS) are potent enhancers of fibroblast growth factor (FGF) - 10 action, we investigated if a C-Xylopyranoside derivative, (C-β-D-xylopyranoside-2-hydroxy-propane, C-Xyloside), could promote DS production by cultured normal human keratinocytes, how this occurs and if C-Xyloside could also stimulate FGF-dependent cell migration and proliferation. C-Xyloside-treated keratinocytes greatly increased secretion of total Sulfated GAGs. Majority of the induced GAG was chondroitin Sulfate/Dermatan Sulfate (CS/DS) of which the major secreted GAG was DS. Cells lacking xylosyltransferase enzymatic activity demonstrated that C-Xyloside was able to stimulate GAG synthesis without addition to core proteins. Consistent with the observed increase in DS, keratinocytes treated with C-Xyloside showed enhanced migration in response to FGF-10 and secreted into their culture media GAGs that promoted FGF-10-dependent cellular proliferation. These results indicate that C-Xyloside may enhance epithelial repair by serving as an initiator of DS synthesis.
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structural and sequence motifs in Dermatan Sulfate for promoting fibroblast growth factor 2 fgf 2 and fgf 7 activity
Journal of Biological Chemistry, 2005Co-Authors: Kristen R Taylor, Jennifer A Rudisill, Richard L GalloAbstract:Glycosaminoglycans have been implicated in the binding and activation of a variety of growth factors, cytokines, and chemokines. In this way, glycosaminoglycans are thought to participate in events such as development and wound repair. In particular, heparin and heparan Sulfate have been well studied, and specific aspects of their structure dictate their participation in a variety of activities. In contrast, although Dermatan Sulfate participates in many of the same biological processes as heparin and heparan Sulfate, the interactions of Dermatan Sulfate have been less well studied. Dermatan Sulfate is abundant in the wound environment and binds and activates growth factors such as fibroblast growth factor-2 (FGF-2) and FGF-7, which are present during the wound repair process. To determine the minimum size and sulfation content of active Dermatan Sulfate oligosaccharides, Dermatan Sulfate was first digested and then separated by size exclusion high pressure liquid chromatography, and the activity to facilitate FGF-2 and FGF-7 was assayed by the cellular proliferation of cell lines expressing FGFR1 or FGFR2 IIIb. The minimum size required for the activation of FGF-2 was an octasaccharide and for FGF-7 a decasaccharide. Active fractions were rich in monoSulfated, primarily 4-O-Sulfated, disaccharides and iduronic acid. Increasing the sulfation to primarily 2/4-O-Sulfated and 2/6-O-Sulfated disaccharides did not increase activity. Cell proliferation decreased or was abolished with higher Sulfated Dermatan Sulfate preparations. This indicated a preference for specific Dermatan Sulfate oligosaccharides capable of promoting FGF-2- and FGF-7-dependent cell proliferation. These data identify critical oligosaccharides that promote specific members of the FGF family that are important for wound repair and angiogenesis.
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Dermatan Sulfate proteoglycan and glycosaminoglycan synthesis is induced in fibroblasts by transfer to a three dimensional extracellular environment
Journal of Biological Chemistry, 2004Co-Authors: Phillip Lee, Janet M Trowbridge, Kristen R Taylor, Vera B Morhenn, Richard L GalloAbstract:Composition and architecture of the extracellular matrix dictate cell behavior. Proteoglycans bind multiple components of the extracellular matrix by serving as important regulators of cell behavior. Given the influence of culture architecture on cell function, we investigated whether switching NIH3T3 fibroblasts from growth on type 1 collagen in monolayer to a collagen gel might influence Dermatan Sulfate expression. Immunofluorescent staining, immunoblot, and Western blot demonstrated an induction in decorin expression in cells switched to collagen gels. This induction was associated with a 40-fold increase in decorin transcript expression determined by quantitative real time PCR. Disaccharide analysis of extracted glycosaminoglycans from collagen gels showed an increase in total glycosaminoglycan and in the ratio of chondroitin Sulfate to heparan Sulfate compared with monolayer culture. The ratio of chondroitin Sulfate to heparan Sulfate likewise increased on syndecan-1 from gel culture. Digestion with chondroitinase B showed that this induced chondroitin Sulfate was Dermatan Sulfate. Syndecan-1 extracted from wounded mouse skin also displayed an increase in Dermatan Sulfate synthesis compared with unwounded skin. Furthermore, glycosaminoglycans from collagen gel culture activated keratinocyte growth factor, whereas glycosaminoglycans from monolayer culture lacked this ability. These findings suggest that regulation of Dermatan Sulfate and Dermatan Sulfate proteoglycan is dependent on extracellular matrix architecture. The ability of collagen gel culture to mimic better the in vivo dermal environment may be due in part to this influence on Dermatan Sulfate and Dermatan Sulfate proteoglycan synthesis.
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Dermatan Sulfate binds and potentiates activity of keratinocyte growth factor fgf 7
Journal of Biological Chemistry, 2002Co-Authors: Janet M Trowbridge, Jennifer A Rudisill, Richard L GalloAbstract:Abstract FGF-7 is induced after injury and induces the proliferation of keratinocytes. Like most members of the FGF family, the activity of FGF-7 is strongly influenced by binding to heparin, but this glycosaminoglycan is absent on keratinocyte cell surfaces and minimally present in the wound environment. In this investigation we compared the relative activity of heparan Sulfate and chondroitin Sulfate B (Dermatan Sulfate), glycosaminoglycans that are present in wounds. A lymphoid cell line (BaF/KGFR) containing the FGF-7 receptor (FGFR2 IIIb) was treated with FGF-7 and with various glycosaminoglycans. FGF-7 did not support cell proliferation in the absence of glycosaminoglycan or with addition of heparan Sulfate or chondroitin Sulfate A/C but did stimulate BaF/KGFR division in the presence of Dermatan Sulfate or highly Sulfated low molecular weight fractions of Dermatan. Dermatan Sulfate also enabled FGF-7-dependent phosphorylation of mitogen-activated protein kinase and promoted binding of radiolabeled FGF-7 to FGFR2 IIIb. In addition, Dermatan Sulfate and FGF-7 stimulated growth of normal keratinocytes in culture. Thus, Dermatan Sulfate, the predominant glycosaminoglycan in skin, is the principle cofactor for FGF-7.
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Dermatan Sulfate new functions from an old glycosaminoglycan
Glycobiology, 2002Co-Authors: Janet M Trowbridge, Richard L GalloAbstract:Glycosaminoglycans constitute a considerable fraction of the glycoconjugates found on cellular membranes and in the extracellular matrix of virtually all mammalian tissues. Their ability to bind and alter protein-protein interactions or enzymatic activity has identified them as important determinants of cellular responsiveness in development, homeostasis, and disease. Although heparan Sulfate tends to be emphasized as the most biologically active glycosaminoglycan, Dermatan Sulfate is a particularly attractive subject for further study because it is expressed in many mammalian tissues and it is the predominant glycan present in skin. Dermatan and Dermatan Sulfate proteoglycans have also been implicated in cardiovascular disease, tumorigenesis, infection, wound repair, and fibrosis. Growing evidence suggests that this glycosaminoglycan, like the better studied heparin and heparan Sulfate, is an important cofactor in a variety of cell behaviors.
Anders Malmström - One of the best experts on this subject based on the ideXlab platform.
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recombinant Dermatan Sulfate is a potent activator of heparin cofactor ii dependent inhibition of thrombin
Glycobiology, 2019Co-Authors: Emil Tykesson, Hanna Thorsson, Anders Malmström, Marco Maccarana, Ulf Ellervik, Gunilla WestergrenthorssonAbstract:The glycosaminoglycan Dermatan Sulfate (DS) is a well-known activator of heparin cofactor II-dependent inactivation of thrombin. In contrast to heparin, Dermatan Sulfate has never been prepared recombinantly from material of non-animal origin. Here we report on the enzymatic synthesis of structurally well-defined DS with high anticoagulant activity. Using a microbial K4 polysaccharide and the recombinant enzymes DS-epimerase 1, Dermatan 4-O-sulfotransferase 1, uronyl 2-O-sulfotransferase and N-acetylgalactosamine 4-Sulfate 6-O-sulfotransferase, several new glycostructures have been prepared, such as a homogenously Sulfated IdoA-GalNAc-4S polymer and its 2-O-, 6-O- and 2,6-O-Sulfated derivatives. Importantly, the recombinant highly 2,4-O-Sulfated DS inhibits thrombin via heparin cofactor II, approximately 20 times better than heparin, enabling manipulation of vascular and extravascular coagulation. The potential of this method can be extended to preparation of specific structures that are of importance for binding and activation of cytokines, and control of inflammation and metastasis, involving extravasation and migration. (Less)
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Dermatan Sulfate epimerase 1 and Dermatan 4 o sulfotransferase 1 form complexes that generate long epimerized 4 o Sulfated blocks
Journal of Biological Chemistry, 2018Co-Authors: Emil Tykesson, Gunilla Westergrenthorsson, Anders Malmström, Ulf Ellervik, Martin A. Thelin, Antti Hassinen, Katarzyna Zielinska, Giacomo Frati, Sakari KellokumpuAbstract:During the biosynthesis of chondroitin/Dermatan Sulfate (CS/DS), a variable fraction of glucuronic acid is converted to iduronic acid through the activities of two epimerases, Dermatan Sulfate epimerases 1 (DS-epi1) and 2 (DS-epi2). Previous in vitro studies indicated that without association with other enzymes, DS-epi1 activity produces structures that have only a few adjacent iduronic acid units. In vivo, concomitant with epimerization, Dermatan 4-O-sulfotransferase 1 (D4ST1) Sulfates the GalNAc adjacent to iduronic acid. This sulfation facilitates DS-epi1 activity and enables the formation of long blocks of Sulfated iduronic acid–containing domains, which can be major components of CS/DS. In this report, we used recombinant enzymes to confirm the concerted action of DS-epi1 and D4ST1. Confocal microscopy revealed that these two enzymes colocalize to the Golgi, and FRET experiments indicated that they physically interact. Furthermore, FRET, immunoprecipitation, and cross-linking experiments also revealed that DS-epi1, DS-epi2, and D4ST1 form homomers and are all part of a hetero-oligomeric complex where D4ST1 directly interacts with DS-epi1, but not with DS-epi2. The cooperation of DS-epi1 with D4ST1 may therefore explain the processive mode of the formation of iduronic acid blocks. In conclusion, the iduronic acid–forming enzymes operate in complexes, similar to other enzymes active in glycosaminoglycan biosynthesis. This knowledge shed light on regulatory mechanisms controlling the biosynthesis of the structurally diverse CS/DS molecule.
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deciphering the mode of action of the processive polysaccharide modifying enzyme Dermatan Sulfate epimerase 1 by hydrogen deuterium exchange mass spectrometry
Chemical Science, 2016Co-Authors: Emil Tykesson, Gunilla Westergrenthorsson, Marco Maccarana, Ulf Ellervik, Lars Malmstrom, Yang Mao, Jinshan Gao, Cheng Lin, Joseph Zaia, Anders MalmströmAbstract:Distinct from template-directed biosynthesis of nucleic acids and proteins, the enzymatic synthesis of heterogeneous polysaccharides is a complex process that is difficult to study using common analytical tools. Therefore, the mode of action and processivity of those enzymes are largely unknown. Dermatan Sulfate epimerase 1 (DS-epi1) is the predominant enzyme during the formation of iduronic acid residues in the glycosaminoglycan Dermatan Sulfate. Using recombinant DS-epi1 as a model enzyme, we describe a tandem mass spectrometry-based method to study the mode of action of polysaccharide processing enzymes. The enzyme action on the substrate was monitored by hydrogen-deuterium exchange mass spectrometry and the sequence information was then fed into mathematical models with two different assumptions of the mode of action for the enzyme: processive reducing end to non-reducing end, and processive non-reducing end to reducing end. Model data was scored by correlation to experimental data and it was found that DS-epi1 attacks its substrate on a random position, followed by a processive mode of modification towards the non-reducing end and that the substrate affinity of the enzyme is negatively affected by each additional epimerization event. It could also be shown that the smallest active substrate was the reducing end uronic acid in a tetrasaccharide and that octasaccharides and longer oligosaccharides were optimal substrates. The method of using tandem mass spectrometry to generate sequence information of the complex enzymatic products in combination with in silico modeling can be potentially applied to study the mode of action of other enzymes involved in polysaccharide biosynthesis.
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biological functions of iduronic acid in chondroitin Dermatan Sulfate
FEBS Journal, 2013Co-Authors: Martin A. Thelin, Emil Tykesson, Marco Maccarana, Ake Oldberg, Barbara Bartolini, Jakob B Axelsson, Renata Gustafsson, Edgar M. Pera, Anders MalmströmAbstract:The presence of iduronic acid in chondroitin/Dermatan Sulfate changes the properties of the polysaccharides because it generates a more flexible chain with increased binding potentials. Iduronic acid in chondroitin/Dermatan Sulfate influences multiple cellular properties, such as migration, proliferation, differentiation, angiogenesis and the regulation of cytokine/growth factor activities. Under pathological conditions such as wound healing, inflammation and cancer, iduronic acid has diverse regulatory functions. Iduronic acid is formed by two epimerases (i.e. Dermatan Sulfate epimerase 1 and 2) that have different tissue distribution and properties. The role of iduronic acid in chondroitin/Dermatan Sulfate is highlighted by the vast changes in connective tissue features in patients with a new type of Ehler–Danlos syndrome: adducted thumb-clubfoot syndrome. Future research aims to understand the roles of the two epimerases and their interplay with the sulfotransferases involved in chondroitin Sulfate/Dermatan Sulfate biosynthesis. Furthermore, a better definition of chondroitin/Dermatan Sulfate functions using different knockout models is needed. In this review, we focus on the two enzymes responsible for iduronic acid formation, as well as the role of iduronic acid in health and disease.
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iduronic acid in chondroitin Dermatan Sulfate biosynthesis and biological function
Journal of Histochemistry and Cytochemistry, 2012Co-Authors: Anders Malmström, Martin A. Thelin, Barbara Bartolini, Benny Pacheco, Marco MaccaranaAbstract:The ability of chondroitin/Dermatan Sulfate (CS/DS) to convey biological information is enriched by the presence of iduronic acid. DS-epimerases 1 and 2 (DS-epi1 and 2), in conjunction with DS-4-O-sulfotransferase 1, are the enzymes responsible for iduronic acid biosynthesis and will be the major focus of this review. CS/DS proteoglycans (CS/DS-PGs) are ubiquitously found in connective tissues, basement membranes, and cell surfaces or are stored intracellularly. Such wide distribution reflects the variety of biological roles in which they are involved, from extracellular matrix organization to regulation of processes such as proliferation, migration, adhesion, and differentiation. They play roles in inflammation, angiogenesis, coagulation, immunity, and wound healing. Such versatility is achieved thanks to their variable composition, both in terms of protein core and the fine structure of the CS/DS chains. Excellent reviews have been published on the collective and individual functions of each CS/DS-PG. This short review presents the biosynthesis and functions of iduronic acid-containing structures, also as revealed by the analysis of the DS-epi1- and 2-deficient mouse models. (J Histochem Cytochem 60: 916-925, 2012) (Less)
Gunilla Westergrenthorsson - One of the best experts on this subject based on the ideXlab platform.
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the structure of human Dermatan Sulfate epimerase 1 emphasizes the importance of c5 epimerization of glucuronic acid in higher organisms
Chemical Science, 2021Co-Authors: Mahmudul Hasan, Gunilla Westergrenthorsson, Hamed Khakzad, Lotta Happonen, Anders Sundin, Johan Unge, U Mueller, Johan Malmstrom, Lars MalmstromAbstract:Dermatan Sulfate epimerase 1 (DS-epi1, EC 5.1.3.19) catalyzes the conversion of D-glucuronic acid to L-iduronic acid on the polymer level, a key step in the biosynthesis of the glycosaminoglycan Dermatan Sulfate. Here, we present the first crystal structure of the catalytic domains of DS-epi1, solved at 2.4 A resolution, as well as a model of the full-length luminal protein obtained by a combination of macromolecular crystallography and targeted cross-linking mass spectrometry. Based on docking studies and molecular dynamics simulations of the protein structure and a chondroitin substrate, we suggest a novel mechanism of DS-epi1, involving a His/double-Tyr motif. Our work uncovers detailed information about the domain architecture, active site, metal-coordinating center and pattern of N-glycosylation of the protein. Additionally, the structure of DS-epi1 reveals a high structural similarity to proteins from several families of bacterial polysaccharide lyases. DS-epi1 is of great importance in a range of diseases, and the structure provides a necessary starting point for design of active site inhibitors.
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recombinant Dermatan Sulfate is a potent activator of heparin cofactor ii dependent inhibition of thrombin
Glycobiology, 2019Co-Authors: Emil Tykesson, Hanna Thorsson, Anders Malmström, Marco Maccarana, Ulf Ellervik, Gunilla WestergrenthorssonAbstract:The glycosaminoglycan Dermatan Sulfate (DS) is a well-known activator of heparin cofactor II-dependent inactivation of thrombin. In contrast to heparin, Dermatan Sulfate has never been prepared recombinantly from material of non-animal origin. Here we report on the enzymatic synthesis of structurally well-defined DS with high anticoagulant activity. Using a microbial K4 polysaccharide and the recombinant enzymes DS-epimerase 1, Dermatan 4-O-sulfotransferase 1, uronyl 2-O-sulfotransferase and N-acetylgalactosamine 4-Sulfate 6-O-sulfotransferase, several new glycostructures have been prepared, such as a homogenously Sulfated IdoA-GalNAc-4S polymer and its 2-O-, 6-O- and 2,6-O-Sulfated derivatives. Importantly, the recombinant highly 2,4-O-Sulfated DS inhibits thrombin via heparin cofactor II, approximately 20 times better than heparin, enabling manipulation of vascular and extravascular coagulation. The potential of this method can be extended to preparation of specific structures that are of importance for binding and activation of cytokines, and control of inflammation and metastasis, involving extravasation and migration. (Less)
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Dermatan Sulfate epimerase 1 and Dermatan 4 o sulfotransferase 1 form complexes that generate long epimerized 4 o Sulfated blocks
Journal of Biological Chemistry, 2018Co-Authors: Emil Tykesson, Gunilla Westergrenthorsson, Anders Malmström, Ulf Ellervik, Martin A. Thelin, Antti Hassinen, Katarzyna Zielinska, Giacomo Frati, Sakari KellokumpuAbstract:During the biosynthesis of chondroitin/Dermatan Sulfate (CS/DS), a variable fraction of glucuronic acid is converted to iduronic acid through the activities of two epimerases, Dermatan Sulfate epimerases 1 (DS-epi1) and 2 (DS-epi2). Previous in vitro studies indicated that without association with other enzymes, DS-epi1 activity produces structures that have only a few adjacent iduronic acid units. In vivo, concomitant with epimerization, Dermatan 4-O-sulfotransferase 1 (D4ST1) Sulfates the GalNAc adjacent to iduronic acid. This sulfation facilitates DS-epi1 activity and enables the formation of long blocks of Sulfated iduronic acid–containing domains, which can be major components of CS/DS. In this report, we used recombinant enzymes to confirm the concerted action of DS-epi1 and D4ST1. Confocal microscopy revealed that these two enzymes colocalize to the Golgi, and FRET experiments indicated that they physically interact. Furthermore, FRET, immunoprecipitation, and cross-linking experiments also revealed that DS-epi1, DS-epi2, and D4ST1 form homomers and are all part of a hetero-oligomeric complex where D4ST1 directly interacts with DS-epi1, but not with DS-epi2. The cooperation of DS-epi1 with D4ST1 may therefore explain the processive mode of the formation of iduronic acid blocks. In conclusion, the iduronic acid–forming enzymes operate in complexes, similar to other enzymes active in glycosaminoglycan biosynthesis. This knowledge shed light on regulatory mechanisms controlling the biosynthesis of the structurally diverse CS/DS molecule.
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deciphering the mode of action of the processive polysaccharide modifying enzyme Dermatan Sulfate epimerase 1 by hydrogen deuterium exchange mass spectrometry
Chemical Science, 2016Co-Authors: Emil Tykesson, Gunilla Westergrenthorsson, Marco Maccarana, Ulf Ellervik, Lars Malmstrom, Yang Mao, Jinshan Gao, Cheng Lin, Joseph Zaia, Anders MalmströmAbstract:Distinct from template-directed biosynthesis of nucleic acids and proteins, the enzymatic synthesis of heterogeneous polysaccharides is a complex process that is difficult to study using common analytical tools. Therefore, the mode of action and processivity of those enzymes are largely unknown. Dermatan Sulfate epimerase 1 (DS-epi1) is the predominant enzyme during the formation of iduronic acid residues in the glycosaminoglycan Dermatan Sulfate. Using recombinant DS-epi1 as a model enzyme, we describe a tandem mass spectrometry-based method to study the mode of action of polysaccharide processing enzymes. The enzyme action on the substrate was monitored by hydrogen-deuterium exchange mass spectrometry and the sequence information was then fed into mathematical models with two different assumptions of the mode of action for the enzyme: processive reducing end to non-reducing end, and processive non-reducing end to reducing end. Model data was scored by correlation to experimental data and it was found that DS-epi1 attacks its substrate on a random position, followed by a processive mode of modification towards the non-reducing end and that the substrate affinity of the enzyme is negatively affected by each additional epimerization event. It could also be shown that the smallest active substrate was the reducing end uronic acid in a tetrasaccharide and that octasaccharides and longer oligosaccharides were optimal substrates. The method of using tandem mass spectrometry to generate sequence information of the complex enzymatic products in combination with in silico modeling can be potentially applied to study the mode of action of other enzymes involved in polysaccharide biosynthesis.
Emil Tykesson - One of the best experts on this subject based on the ideXlab platform.
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recombinant Dermatan Sulfate is a potent activator of heparin cofactor ii dependent inhibition of thrombin
Glycobiology, 2019Co-Authors: Emil Tykesson, Hanna Thorsson, Anders Malmström, Marco Maccarana, Ulf Ellervik, Gunilla WestergrenthorssonAbstract:The glycosaminoglycan Dermatan Sulfate (DS) is a well-known activator of heparin cofactor II-dependent inactivation of thrombin. In contrast to heparin, Dermatan Sulfate has never been prepared recombinantly from material of non-animal origin. Here we report on the enzymatic synthesis of structurally well-defined DS with high anticoagulant activity. Using a microbial K4 polysaccharide and the recombinant enzymes DS-epimerase 1, Dermatan 4-O-sulfotransferase 1, uronyl 2-O-sulfotransferase and N-acetylgalactosamine 4-Sulfate 6-O-sulfotransferase, several new glycostructures have been prepared, such as a homogenously Sulfated IdoA-GalNAc-4S polymer and its 2-O-, 6-O- and 2,6-O-Sulfated derivatives. Importantly, the recombinant highly 2,4-O-Sulfated DS inhibits thrombin via heparin cofactor II, approximately 20 times better than heparin, enabling manipulation of vascular and extravascular coagulation. The potential of this method can be extended to preparation of specific structures that are of importance for binding and activation of cytokines, and control of inflammation and metastasis, involving extravasation and migration. (Less)
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Dermatan Sulfate epimerase 1 and Dermatan 4 o sulfotransferase 1 form complexes that generate long epimerized 4 o Sulfated blocks
Journal of Biological Chemistry, 2018Co-Authors: Emil Tykesson, Gunilla Westergrenthorsson, Anders Malmström, Ulf Ellervik, Martin A. Thelin, Antti Hassinen, Katarzyna Zielinska, Giacomo Frati, Sakari KellokumpuAbstract:During the biosynthesis of chondroitin/Dermatan Sulfate (CS/DS), a variable fraction of glucuronic acid is converted to iduronic acid through the activities of two epimerases, Dermatan Sulfate epimerases 1 (DS-epi1) and 2 (DS-epi2). Previous in vitro studies indicated that without association with other enzymes, DS-epi1 activity produces structures that have only a few adjacent iduronic acid units. In vivo, concomitant with epimerization, Dermatan 4-O-sulfotransferase 1 (D4ST1) Sulfates the GalNAc adjacent to iduronic acid. This sulfation facilitates DS-epi1 activity and enables the formation of long blocks of Sulfated iduronic acid–containing domains, which can be major components of CS/DS. In this report, we used recombinant enzymes to confirm the concerted action of DS-epi1 and D4ST1. Confocal microscopy revealed that these two enzymes colocalize to the Golgi, and FRET experiments indicated that they physically interact. Furthermore, FRET, immunoprecipitation, and cross-linking experiments also revealed that DS-epi1, DS-epi2, and D4ST1 form homomers and are all part of a hetero-oligomeric complex where D4ST1 directly interacts with DS-epi1, but not with DS-epi2. The cooperation of DS-epi1 with D4ST1 may therefore explain the processive mode of the formation of iduronic acid blocks. In conclusion, the iduronic acid–forming enzymes operate in complexes, similar to other enzymes active in glycosaminoglycan biosynthesis. This knowledge shed light on regulatory mechanisms controlling the biosynthesis of the structurally diverse CS/DS molecule.
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deciphering the mode of action of the processive polysaccharide modifying enzyme Dermatan Sulfate epimerase 1 by hydrogen deuterium exchange mass spectrometry
Chemical Science, 2016Co-Authors: Emil Tykesson, Gunilla Westergrenthorsson, Marco Maccarana, Ulf Ellervik, Lars Malmstrom, Yang Mao, Jinshan Gao, Cheng Lin, Joseph Zaia, Anders MalmströmAbstract:Distinct from template-directed biosynthesis of nucleic acids and proteins, the enzymatic synthesis of heterogeneous polysaccharides is a complex process that is difficult to study using common analytical tools. Therefore, the mode of action and processivity of those enzymes are largely unknown. Dermatan Sulfate epimerase 1 (DS-epi1) is the predominant enzyme during the formation of iduronic acid residues in the glycosaminoglycan Dermatan Sulfate. Using recombinant DS-epi1 as a model enzyme, we describe a tandem mass spectrometry-based method to study the mode of action of polysaccharide processing enzymes. The enzyme action on the substrate was monitored by hydrogen-deuterium exchange mass spectrometry and the sequence information was then fed into mathematical models with two different assumptions of the mode of action for the enzyme: processive reducing end to non-reducing end, and processive non-reducing end to reducing end. Model data was scored by correlation to experimental data and it was found that DS-epi1 attacks its substrate on a random position, followed by a processive mode of modification towards the non-reducing end and that the substrate affinity of the enzyme is negatively affected by each additional epimerization event. It could also be shown that the smallest active substrate was the reducing end uronic acid in a tetrasaccharide and that octasaccharides and longer oligosaccharides were optimal substrates. The method of using tandem mass spectrometry to generate sequence information of the complex enzymatic products in combination with in silico modeling can be potentially applied to study the mode of action of other enzymes involved in polysaccharide biosynthesis.
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biological functions of iduronic acid in chondroitin Dermatan Sulfate
FEBS Journal, 2013Co-Authors: Martin A. Thelin, Emil Tykesson, Marco Maccarana, Ake Oldberg, Barbara Bartolini, Jakob B Axelsson, Renata Gustafsson, Edgar M. Pera, Anders MalmströmAbstract:The presence of iduronic acid in chondroitin/Dermatan Sulfate changes the properties of the polysaccharides because it generates a more flexible chain with increased binding potentials. Iduronic acid in chondroitin/Dermatan Sulfate influences multiple cellular properties, such as migration, proliferation, differentiation, angiogenesis and the regulation of cytokine/growth factor activities. Under pathological conditions such as wound healing, inflammation and cancer, iduronic acid has diverse regulatory functions. Iduronic acid is formed by two epimerases (i.e. Dermatan Sulfate epimerase 1 and 2) that have different tissue distribution and properties. The role of iduronic acid in chondroitin/Dermatan Sulfate is highlighted by the vast changes in connective tissue features in patients with a new type of Ehler–Danlos syndrome: adducted thumb-clubfoot syndrome. Future research aims to understand the roles of the two epimerases and their interplay with the sulfotransferases involved in chondroitin Sulfate/Dermatan Sulfate biosynthesis. Furthermore, a better definition of chondroitin/Dermatan Sulfate functions using different knockout models is needed. In this review, we focus on the two enzymes responsible for iduronic acid formation, as well as the role of iduronic acid in health and disease.