Structural analysis of chondroitin sulfate from Scyliorhinus canicula: a useful source of this polysaccharide.

Chondroitin sulfate (CS), a constituent of proteoglycans, is a key component of the connective tissues and it is widely used as a precautionary drug for joint diseases; for this reason, the increased demand of this polysaccharide has posed the problem to identify new and secure sources of this product. In this context, CS from the cartilage of the lesser spotted dogfish (Scyliorhinus canicula, a cartilaginous fish) was isolated and investigated through chemical and spectroscopical techniques. The structural elucidation was performed on the entire polysaccharide and confirmed analyzing the products obtained via ABC lyase treatment. As a result, its compositional analysis disclosed the occurrence of CS-A, CS-C, CS-D, and CS-0S motifs in the ratio of 41, 32, 19.8, and 8.2%, respectively. Additionally, two different glycopeptides were isolated and characterized via NMR, providing information on the linkage oligosaccharide region joining the glycosaminoglycan chain to the core protein. Therefore, chondroitin sulfate from Scyliorhinus canicula appears very similar to that isolated from shark, a cartilaginous and taxonomically related fish, with the main difference residing in the major percentage of the CS-A motif. In the light of the results obtained, Scyliorhinus canicula chondroitin sulfate possesses a chemical structure compatible for the formulation of commercial and pharmaceutical products.

[1]  N. Volpi Advances in chondroitin sulfate analysis: application in physiological and pathological States of connective tissue and during pharmacological treatment of osteoarthritis. , 2006, Current pharmaceutical design.

[2]  G. Torri,et al.  Complex glycosaminoglycans: profiling substitution patterns by two-dimensional nuclear magnetic resonance spectroscopy. , 2005, Analytical biochemistry.

[3]  A. Mucci,et al.  1H and 13C nuclear magnetic resonance identification and characterization of components of chondroitin sulfates of various origin , 2000 .

[4]  N. Volpi Disaccharide mapping of chondroitin sulfate of different origins by high-performance capillary electrophoresis and high-performance liquid chromatography , 2004 .

[5]  Robert J Linhardt,et al.  Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides* , 2004, Journal of Biological Chemistry.

[6]  R. Midura,et al.  Isolation and characterization of proteoglycans. , 1994, Methods in enzymology.

[7]  P. Janvier,et al.  The complete nucleotide sequence of the mitochondrial DNA of the dogfish, Scyliorhinus canicula. , 1998, Genetics.

[8]  B. Mulloy,et al.  Structure of a fucose-branched chondroitin sulfate from sea cucumber. Evidence for the presence of 3-O-sulfo-beta-D-glucuronosyl residues. , 1991, The Journal of biological chemistry.

[9]  Rahul Raman,et al.  Glycomics approach to structure-function relationships of glycosaminoglycans. , 2006, Annual review of biomedical engineering.

[10]  O. Holst Bacterial toxins : methods and protocols , 2000 .

[11]  V. Hascall,et al.  Cell surface heparan sulfate proteoglycans. , 1992, The Journal of biological chemistry.

[12]  D. Heinegård,et al.  [17] Isolation and characterization of proteoglycans , 1987 .

[13]  N. Volpi On-line HPLC/ESI-MS separation and characterization of hyaluronan oligosaccharides from 2-mers to 40-mers. , 2007, Analytical chemistry.

[14]  J. Vliegenthart,et al.  Structural studies on sulfated glycopeptides from the carbohydrate-protein linkage region of chondroitin 4-sulfate proteoglycans of swarm rat chondrosarcoma. Demonstration of the structure Gal(4-O-sulfate)β1-3Galβ1-4Xylβ1-O-Ser , 1988 .

[15]  P. de Waard,et al.  Structural studies on sulfated glycopeptides from the carbohydrate-protein linkage region of chondroitin 4-sulfate proteoglycans of swarm rat chondrosarcoma. Demonstration of the structure Gal(4-O-sulfate)beta 1-3Gal beta 1-4XYL beta 1-O-Ser. , 1988, The Journal of biological chemistry.

[16]  K. Yoshida,et al.  One- and two-dimensional 1H-NMR characterization of two series of sulfated disaccharides prepared from chondroitin sulfate and heparan sulfate/heparin by bacterial eliminase digestion. , 1992, Journal of biochemistry.

[17]  Zhenqing Zhang,et al.  Mass spectrometry for the characterization of unsulfated chondroitin oligosaccharides from 2-mers to 16-mers. Comparison with hyaluronic acid oligomers. , 2008, Rapid communications in mass spectrometry : RCM.

[18]  Kazuyuki Sugahara,et al.  Recent Advances in the Structural Study of Functional Chondroitin Sulfate and Dermatan Sulfate in Health and Disease , 2008, Connective tissue research.

[19]  H. Kitagawa,et al.  Novel Sulfated Oligosaccharides Containing 3-O-Sulfated Glucuronic Acid from King Crab Cartilage Chondroitin Sulfate K , 1996, The Journal of Biological Chemistry.

[20]  T. Hardingham,et al.  Proteoglycans of articular cartilage: changes in aging and in joint disease. , 1990, Seminars in arthritis and rheumatism.

[21]  Robert Langer,et al.  Oversulfated chondroitin sulfate is a contaminant in heparin associated with adverse clinical events , 2008, Nature Biotechnology.

[22]  R. Iozzo Matrix proteoglycans: from molecular design to cellular function. , 1998, Annual review of biochemistry.

[23]  K. A. Taylor,et al.  A modification of the phenol/sulfuric acid assay for total carbohydrates giving more comparable absorbances , 1995 .

[24]  O. Habuchi,et al.  Glucose branches in chondroitin sulfates from squid cartilage. , 1977, The Journal of biological chemistry.