Novel Chondroitin Sulfate Oligosaccharide Motifs as Biomarkers: Insights into Their Involvement in Brain Development
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Chondroitin sulfate (CS) chains regulate the development of the central nervous system in vertebrates. They are the linear polysaccharide side chains of proteoglycans that consist of various sulfated repeating disaccharides, [–4GlcUAβ1–3GalNAcβ1–]n, in which GlcUA and GalNAc represent d-glucuronic acid and N-acetyl-d-galactosamine, respectively. CS chains containing disaccharide designated D-units, GlcUA(2-O-sulfate)–GalNAc(6-O-sulfate), play a role in brain development by interacting with the neurotrophic factor, pleiotrophin, which generates signaling. A series of octasaccharide sequences containing at least one D-unit, were previously isolated from embryonic pig brains using a pleiotrophin-immobilized affinity column. These similar yet distinct sugar sequences were termed “wobble oligosaccharide motifs” on account of their structural flexibility. Oligosaccharides containing D-units were recently isolated from shark fin cartilage to obtain more detailed structural information on the CS chains containing D-units that are involved in brain development. Seven novel hexasaccharide sequences, in addition to three previously reported sequences, were isolated after exhaustive digestion of the CS chains with the chondroitin lyase AC-I, which cannot act on galactosaminidic linkages adjacent to the D-units. Of the epitopes involved in cerebellar development in mammals, the novel epitope sequences ∆A-D-A, ∆A-D-D, and ∆A-B-D were identified in binding studies using an anti-CS monoclonal antibody, with the symbol ∆ referring to the 4,5-unsaturated bond of GlcUA residues generated by the lyase, while A, B, and C represented GlcUA-GalNAc(4-O-sulfate), GlcUA(2-O-sulfate)-GalNAc(4-O-sulfate), and GlcUA-GalNAc(6-O-sulfate), respectively. The significance of multiple CS oligosaccharide sequences in the brain and electrostatic potential distribution at their molecular surface during ligand binding have been discussed.
KeywordsChondroitin sulfate Dermatan sulfate Sugar sequences Sugar biomarkers Brain development
Chondroitin 4-O-sulfotransferase 1
Chondroitin 4-O-sulfotransferase 2
This work was supported in part by a Grant-in-Aid for Challenging Exploratory Research 25670018 and the Japan–Thailand Research Cooperative Program from the Japan Society for the Promotion of Science (JSPS), and a Grant in-Aid for Scientific Research on Innovative Areas 24110501 from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT).
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