title: Novel 70-kDa chondroitin sulfate/dermatan sulfate hybrid chains with a unique heterogeneous sulfation pattern from shark skin, which exhibit neuritogenic activity and binding activities for growth factors and neurotrophic factors.
abstract: Chondroitin sulfate (CS) and dermatan sulfate (DS) hybrid chains of proteoglycans are critical in growth factor binding, neuritogenesis, and brain development. Here we isolated CS/DS hybrid chains from shark skin aiming to develop therapeutic agents. Digestion with various chondroitinases showed that both GlcUA- and IdoUA-containing disaccharides are scattered along the polysaccharide chains with an unusually large average molecular mass of 70 kDa. The CS/DS chains were separated into major (80%) and minor (20%) fractions by anion-exchange chromatography. Both fractions had relatively low degrees of sulfation (sulfate/disaccharide molar ratio=1.17 versus 0.87), showing a unique feature compared with the marine CS and DS isolated to date, most of which are oversulfated. They were highly heterogeneous and characterized by multiple disaccharides including GlcUA-GalNAc, GlcUA-GalNAc(6S), GlcUA-GalNAc(4S), IdoUA-GalNAc(4S), GlcUA-GalNAc(4S,6S), IdoUA-GalNAc(4S,6S), GlcUA(2S)-GalNAc(6S), and/or IdoUA(2S)-GalNAc(6S), IdoUA(2S)-GalNAc(4S) and novel GlcUA(2S)-GalNAc(4S), where 2S, 4S, and 6S represent 2-O-, 4-O- and 6-O-sulfate, respectively. The CS/DS chains bound two neurotrophic factors and various growth factors expressed in the brain with high affinity as evaluated for the major fraction by kinetic analysis using a surface plasmon resonance detector, and also promoted the outgrowth of neurites of both an axonic and a dendritic nature. The neuritogenic activity was abolished completely by digestion with chondroitinase ABC, AC-I, or B, suggesting the importance of both GlcUA- and IdoUA-containing moieties. It also showed anti-heparin cofactor II activity comparable to that exhibited by DS from porcine skin. Thus, by virtue of its unique structure and biological activities, DS will find a potential use in therapeutics.
authors.affiliation: Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan.
authors.forename: Chilkunda D
authors.initials: CD
authors.lastname: Nandini
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Itoh
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Sugahara
mesh_terms: D000818:Animals; D019921:Biotinylation; D049109:Cell Proliferation; D019765:Chondroitin ABC Lyase / Q000737:chemistry; D002809:Chondroitin Sulfates / Q000737:chemistry*; D002810:Chondroitinases and Chondroitin Lyases / Q000737:chemistry; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003871:Dermatan Sulfate / Q000737:chemistry*; D004187:Disaccharides / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004848:Epithelium / Q000378:metabolism; D006025:Glycosaminoglycans / Q000737:chemistry; D006133:Growth Substances / Q000378:metabolism*; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000737:chemistry; D007700:Kinetics; D009414:Nerve Growth Factors / Q000737:chemistry*; D009474:Neurons / Q000378:metabolism; D009608:Nitrous Acid / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D011485:Protein Binding; D011516:Prothrombin / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012754:Sharks; D020349:Surface Plasmon Resonance; D013552:Swine; D013997:Time Factors
chemical_list: D004187:Disaccharides; D006025:Glycosaminoglycans; D006133:Growth Substances; D009414:Nerve Growth Factors; D010455:Peptides; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; D003871:Dermatan Sulfate; D011516:Prothrombin; C025337:Factor IIa; D002809:Chondroitin Sulfates; D006821:Hyaluronoglucosaminidase; D002810:Chondroitinases and Chondroitin Lyases; D019765:Chondroitin ABC Lyase; D009608:Nitrous Acid