The Experts below are selected from a list of 1452 Experts worldwide ranked by ideXlab platform
Shigeyuki Fukui - One of the best experts on this subject based on the ideXlab platform.
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detection of oligosaccharide ligands for hepatocyte growth factor scatter factor hgf sf keratinocyte growth factor kgf fgf 7 rantes and heparin cofactor ii by neoglycolipid microarrays of glycosaminoglycan derived oligosaccharide fragments
Glycoconjugate Journal, 2006Co-Authors: Keiko Yamaguchi, Hirotoshi Tamaki, Shigeyuki FukuiAbstract:Neoglycolipid technology is eminently adaptable for microarray design for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Dermatan sulfate (DS) is known to play an important role because of its ability to bind growth factors as well as chemokines and to modulate their biological activities during inflammation and response to injury. We prepared various Iduronic Acid-rich fragments from DS by complete digestion with chondroitinase ACI, and investigated whether the DS-binding proteins, such as HGF/SF, RANTES, KGF/FGF-7 and HCII, can detect their oligosaccharide ligands in a neoglycolipid microarray. First, a comparison of the intensity of binding signals obtained from chondroitin oligosaccharides with those of heparin oligosaccharides showed that our microarray system is feasible not only to single-out the oligosaccharide ligands, but also to detect the difference between an intrinsic interaction unrelated only to electrostatic interaction and non-specific electrostatic interaction. Second, HGF/SF, KGF/FGF-7 and HCII showed preferential binding to Iduronic Acid-rich fragments of DS oligosaccharides that are greater than 8-mers in lengths. In contrast, RANTES binding seemed to depend only on the negative charges; their binding intensity towards the DS oligosaccharides was somewhat stronger than the binding of HGF/SF, KGF/FGF-7 and HCII. Third, the use of polyvinylpyrrolidone-40 (PVP-40), ovalbumin (OV) and Tween 20 in place of BSA as a blotting agent was useful in these glycosaminoglycan dependent reactions to minimize background due to non-specific interactions.
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detection of oligosaccharide ligands for hepatocyte growth factor scatter factor hgf sf keratinocyte growth factor kgf fgf 7 rantes and heparin cofactor ii by neoglycolipid microarrays of glycosaminoglycan derived oligosaccharide fragments
Glycoconjugate Journal, 2006Co-Authors: Keiko Yamaguchi, Hirotoshi Tamaki, Shigeyuki FukuiAbstract:Neoglycolipid technology is eminently adaptable for microarray design for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Dermatan sulfate (DS) is known to play an important role because of its ability to bind growth factors as well as chemokines and to modulate their biological activities during inflammation and response to injury. We prepared various Iduronic Acid-rich fragments from DS by complete digestion with chondroitinase ACI, and investigated whether the DS-binding proteins, such as HGF/SF, RANTES, KGF/FGF-7 and HCII, can detect their oligosaccharide ligands in a neoglycolipid microarray. First, a comparison of the intensity of binding signals obtained from chondroitin oligosaccharides with those of heparin oligosaccharides showed that our microarray system is feasible not only to single-out the oligosaccharide ligands, but also to detect the difference between an intrinsic interaction unrelated only to electrostatic interaction and non-specific electrostatic interaction. Second, HGF/SF, KGF/FGF-7 and HCII showed preferential binding to Iduronic Acid-rich fragments of DS oligosaccharides that are greater than 8-mers in lengths. In contrast, RANTES binding seemed to depend only on the negative charges; their binding intensity towards the DS oligosaccharides was somewhat stronger than the binding of HGF/SF, KGF/FGF-7 and HCII. Third, the use of polyvinylpyrrolidone-40 (PVP-40), ovalbumin (OV) and Tween 20 in place of BSA as a blotting agent was useful in these glycosaminoglycan dependent reactions to minimize background due to non-specific interactions.
Keiko Yamaguchi - One of the best experts on this subject based on the ideXlab platform.
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detection of oligosaccharide ligands for hepatocyte growth factor scatter factor hgf sf keratinocyte growth factor kgf fgf 7 rantes and heparin cofactor ii by neoglycolipid microarrays of glycosaminoglycan derived oligosaccharide fragments
Glycoconjugate Journal, 2006Co-Authors: Keiko Yamaguchi, Hirotoshi Tamaki, Shigeyuki FukuiAbstract:Neoglycolipid technology is eminently adaptable for microarray design for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Dermatan sulfate (DS) is known to play an important role because of its ability to bind growth factors as well as chemokines and to modulate their biological activities during inflammation and response to injury. We prepared various Iduronic Acid-rich fragments from DS by complete digestion with chondroitinase ACI, and investigated whether the DS-binding proteins, such as HGF/SF, RANTES, KGF/FGF-7 and HCII, can detect their oligosaccharide ligands in a neoglycolipid microarray. First, a comparison of the intensity of binding signals obtained from chondroitin oligosaccharides with those of heparin oligosaccharides showed that our microarray system is feasible not only to single-out the oligosaccharide ligands, but also to detect the difference between an intrinsic interaction unrelated only to electrostatic interaction and non-specific electrostatic interaction. Second, HGF/SF, KGF/FGF-7 and HCII showed preferential binding to Iduronic Acid-rich fragments of DS oligosaccharides that are greater than 8-mers in lengths. In contrast, RANTES binding seemed to depend only on the negative charges; their binding intensity towards the DS oligosaccharides was somewhat stronger than the binding of HGF/SF, KGF/FGF-7 and HCII. Third, the use of polyvinylpyrrolidone-40 (PVP-40), ovalbumin (OV) and Tween 20 in place of BSA as a blotting agent was useful in these glycosaminoglycan dependent reactions to minimize background due to non-specific interactions.
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detection of oligosaccharide ligands for hepatocyte growth factor scatter factor hgf sf keratinocyte growth factor kgf fgf 7 rantes and heparin cofactor ii by neoglycolipid microarrays of glycosaminoglycan derived oligosaccharide fragments
Glycoconjugate Journal, 2006Co-Authors: Keiko Yamaguchi, Hirotoshi Tamaki, Shigeyuki FukuiAbstract:Neoglycolipid technology is eminently adaptable for microarray design for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Dermatan sulfate (DS) is known to play an important role because of its ability to bind growth factors as well as chemokines and to modulate their biological activities during inflammation and response to injury. We prepared various Iduronic Acid-rich fragments from DS by complete digestion with chondroitinase ACI, and investigated whether the DS-binding proteins, such as HGF/SF, RANTES, KGF/FGF-7 and HCII, can detect their oligosaccharide ligands in a neoglycolipid microarray. First, a comparison of the intensity of binding signals obtained from chondroitin oligosaccharides with those of heparin oligosaccharides showed that our microarray system is feasible not only to single-out the oligosaccharide ligands, but also to detect the difference between an intrinsic interaction unrelated only to electrostatic interaction and non-specific electrostatic interaction. Second, HGF/SF, KGF/FGF-7 and HCII showed preferential binding to Iduronic Acid-rich fragments of DS oligosaccharides that are greater than 8-mers in lengths. In contrast, RANTES binding seemed to depend only on the negative charges; their binding intensity towards the DS oligosaccharides was somewhat stronger than the binding of HGF/SF, KGF/FGF-7 and HCII. Third, the use of polyvinylpyrrolidone-40 (PVP-40), ovalbumin (OV) and Tween 20 in place of BSA as a blotting agent was useful in these glycosaminoglycan dependent reactions to minimize background due to non-specific interactions.
Anders Malmström - One of the best experts on this subject based on the ideXlab platform.
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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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DKO mice are DS-free.
2015Co-Authors: Xanthi N. Stachtea, Emil Tykesson, Anders Malmström, Toin H Van Kuppevelt, Ricardo Feinstein, Rogier M. Reijmers, Marco MaccaranaAbstract:Two-day-old pups were metabolically labeled with 35S sulfate. Skin decorin CS/DS was extracted and purified (A-C). A) CS/DS chains were analyzed on a size-permeation Superose 6 column. The HS degraded products, which were proportionally increased in the DKO mice, derive from the HS-PGs which co-eluted with decorin in the size permeation column used for PGs separation (see “Material and methods”). B) Split products obtained after chondroitinase B treatment were size separated on a Superdex Peptide column and C) the Iduronic Acid content was calculated. The Iduronic Acid analysis was conducted in duplicate samples. D) Skin PGs were extracted from E19.5 embryos of different genotypes. Decorin was stained before and after chondroitinase ABC and B treatment.
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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, Benny Pacheco, Martin A. Thelin, Barbara Bartolini, 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)
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dermatan sulfate epimerase 1 deficient mice have reduced content and changed distribution of Iduronic Acids in dermatan sulfate and an altered collagen structure in skin
Molecular and Cellular Biology, 2009Co-Authors: Marco Maccarana, Sebastian Kalamajski, M Kongsgaard, Peter S Magnusson, Ake Oldberg, Anders MalmströmAbstract:Dermatan sulfate epimerase 1 (DS-epi1) and DS-epi2 convert glucuronic Acid to Iduronic Acid in chondroitin/dermatan sulfate biosynthesis. Here we report on the generation of DS-epi1-null mice and the resulting alterations in the chondroitin/dermatan polysaccharide chains. The numbers of long blocks of adjacent Iduronic Acids are greatly decreased in skin decorin and biglycan chondroitin/dermatan sulfate, along with a parallel decrease in Iduronic-2-O-sulfated-galactosamine-4-O-sulfated structures. Both Iduronic Acid blocks and Iduronic Acids surrounded by glucuronic Acids are also decreased in versican-derived chains. DS-epi1-deficient mice are smaller than their wild-type littermates but otherwise have no gross macroscopic alterations. The lack of DS-epi1 affects the chondroitin/dermatan sulfate in many proteoglycans, and the consequences for skin collagen structure were initially analyzed. We found that the skin collagen architecture was altered, and electron microscopy showed that the DS-epi1-null fibrils have a larger diameter than the wild-type fibrils. The altered chondroitin/dermatan sulfate chains carried by decorin in skin are likely to affect collagen fibril formation and reduce the tensile strength of DS-epi1-null skin.
Marco Maccarana - One of the best experts on this subject based on the ideXlab platform.
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aberrant neural crest development causes craniofacial and other malformations in an animal model of musculocontractural ehlers danlos syndrome
Journal of Rare Diseases Research & Treatment, 2016Co-Authors: Edgar M. Pera, Nadege Gouignard, Marco MaccaranaAbstract:Musculocontractural Ehlers-Danlos syndrome (MC-EDS) is a rare recessive disorder that is characterized by connective tissue fragility, distinct craniofacial features and congenital malformations. MC-EDS patients have defects in the enzymes dermatan sulfate epimerase-1 and dermatan 4-O-sulfotransferase-1, which are involved in the biosynthesis of Iduronic Acid in the chondroitin sulfate/dermatan sulfate (CS/DS) chains of proteoglycans (PGs). While the connective tissue defect is a result of disturbed collagen fibril assembly based on a decreased Iduronic Acid content of interacting CS/DS-PGs, the cause of the developmental malformations in MC-EDS is not well understood. This review focuses on a new role of CS/DS-PGs in the development of multipotent and highly migratory neural crest (NC) cells in the Xenopus embryo model of MC-EDS. Single Iduronic Acid residues in CS/DS-PGs are involved in the formation of NC-derived craniofacial structures by facilitating the migration and adhesion of NC cells to fibronectin. Our results suggest a defect in NC development as cause of the craniofacial and other congenital anomalies in MC-EDS patients, which might contribute to an improved diagnosis and etiology-based therapy. (Less)
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Maccarana M. Iduronic Acid in chondroitin/dermatan sulfate: biosynthesis and biological function
2016Co-Authors: Barbara Bartolini, Martin A. Thelin, Lena Svensson, Giancarlo Ghiselli, Marco MaccaranaAbstract:Aortic smooth muscle cells produce chondroitin/dermatan sulfate (CS/DS) proteoglycans that regulate extracellular matrix organization and cell behavior in normal and pathological conditions. A unique feature of CS/DS proteoglycans is the presence of Iduronic Acid (IdoA), catalyzed by two DS epimerases. Functional ablation of DS-epi1, the main epimerase in these cells, resulted in a major reduction of IdoA both on cell surface and in secreted CS/DS proteoglycans. Downregulation of IdoA led to delayed ability to re-populate wounded areas due to loss of directional persistence of migration. DS-epi12/2 aortic smooth muscle cells, however, had not lost the general property of migration showing even increased speed of movement compared to wild type cells. Where the cell membrane adheres to the substratum, stress fibers were denser whereas focal adhesion sites were fewer. Total cellular expression of focal adhesion kinase (FAK) and phospho-FAK (pFAK) was decreased in mutant cells compared to control cells. As many pathological conditions are dependent on migration, modulation of IdoA content may point to therapeutic strategies for diseases such as cancer and atherosclerosis
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DKO mice are DS-free.
2015Co-Authors: Xanthi N. Stachtea, Emil Tykesson, Anders Malmström, Toin H Van Kuppevelt, Ricardo Feinstein, Rogier M. Reijmers, Marco MaccaranaAbstract:Two-day-old pups were metabolically labeled with 35S sulfate. Skin decorin CS/DS was extracted and purified (A-C). A) CS/DS chains were analyzed on a size-permeation Superose 6 column. The HS degraded products, which were proportionally increased in the DKO mice, derive from the HS-PGs which co-eluted with decorin in the size permeation column used for PGs separation (see “Material and methods”). B) Split products obtained after chondroitinase B treatment were size separated on a Superdex Peptide column and C) the Iduronic Acid content was calculated. The Iduronic Acid analysis was conducted in duplicate samples. D) Skin PGs were extracted from E19.5 embryos of different genotypes. Decorin was stained before and after chondroitinase ABC and B treatment.
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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, Benny Pacheco, Martin A. Thelin, Barbara Bartolini, 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)
Hirotoshi Tamaki - One of the best experts on this subject based on the ideXlab platform.
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detection of oligosaccharide ligands for hepatocyte growth factor scatter factor hgf sf keratinocyte growth factor kgf fgf 7 rantes and heparin cofactor ii by neoglycolipid microarrays of glycosaminoglycan derived oligosaccharide fragments
Glycoconjugate Journal, 2006Co-Authors: Keiko Yamaguchi, Hirotoshi Tamaki, Shigeyuki FukuiAbstract:Neoglycolipid technology is eminently adaptable for microarray design for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Dermatan sulfate (DS) is known to play an important role because of its ability to bind growth factors as well as chemokines and to modulate their biological activities during inflammation and response to injury. We prepared various Iduronic Acid-rich fragments from DS by complete digestion with chondroitinase ACI, and investigated whether the DS-binding proteins, such as HGF/SF, RANTES, KGF/FGF-7 and HCII, can detect their oligosaccharide ligands in a neoglycolipid microarray. First, a comparison of the intensity of binding signals obtained from chondroitin oligosaccharides with those of heparin oligosaccharides showed that our microarray system is feasible not only to single-out the oligosaccharide ligands, but also to detect the difference between an intrinsic interaction unrelated only to electrostatic interaction and non-specific electrostatic interaction. Second, HGF/SF, KGF/FGF-7 and HCII showed preferential binding to Iduronic Acid-rich fragments of DS oligosaccharides that are greater than 8-mers in lengths. In contrast, RANTES binding seemed to depend only on the negative charges; their binding intensity towards the DS oligosaccharides was somewhat stronger than the binding of HGF/SF, KGF/FGF-7 and HCII. Third, the use of polyvinylpyrrolidone-40 (PVP-40), ovalbumin (OV) and Tween 20 in place of BSA as a blotting agent was useful in these glycosaminoglycan dependent reactions to minimize background due to non-specific interactions.
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detection of oligosaccharide ligands for hepatocyte growth factor scatter factor hgf sf keratinocyte growth factor kgf fgf 7 rantes and heparin cofactor ii by neoglycolipid microarrays of glycosaminoglycan derived oligosaccharide fragments
Glycoconjugate Journal, 2006Co-Authors: Keiko Yamaguchi, Hirotoshi Tamaki, Shigeyuki FukuiAbstract:Neoglycolipid technology is eminently adaptable for microarray design for high-throughput detection and specificity assignments of carbohydrate-protein interactions. Dermatan sulfate (DS) is known to play an important role because of its ability to bind growth factors as well as chemokines and to modulate their biological activities during inflammation and response to injury. We prepared various Iduronic Acid-rich fragments from DS by complete digestion with chondroitinase ACI, and investigated whether the DS-binding proteins, such as HGF/SF, RANTES, KGF/FGF-7 and HCII, can detect their oligosaccharide ligands in a neoglycolipid microarray. First, a comparison of the intensity of binding signals obtained from chondroitin oligosaccharides with those of heparin oligosaccharides showed that our microarray system is feasible not only to single-out the oligosaccharide ligands, but also to detect the difference between an intrinsic interaction unrelated only to electrostatic interaction and non-specific electrostatic interaction. Second, HGF/SF, KGF/FGF-7 and HCII showed preferential binding to Iduronic Acid-rich fragments of DS oligosaccharides that are greater than 8-mers in lengths. In contrast, RANTES binding seemed to depend only on the negative charges; their binding intensity towards the DS oligosaccharides was somewhat stronger than the binding of HGF/SF, KGF/FGF-7 and HCII. Third, the use of polyvinylpyrrolidone-40 (PVP-40), ovalbumin (OV) and Tween 20 in place of BSA as a blotting agent was useful in these glycosaminoglycan dependent reactions to minimize background due to non-specific interactions.
