The DSD-1 carbohydrate epitope depends on sulfation, correlates with chondroitin sulfate D motifs, and is sufficient to promote neurite outgrowth

被引:168
作者
Clement, AM
Nadanaka, S
Masayama, K
Mandl, C
Sugahara, K
Faissner, A
机构
[1] Univ Heidelberg, Dept Neurobiol, D-69120 Heidelberg, Germany
[2] Kobe Pharmaceut Univ, Dept Biochem, Kobe, Hyogo 6588558, Japan
[3] CNRS, Ctr Neurochim, Lab Neurobiol Dev & Regenerat, UPR 1352, F-67084 Strasbourg, France
[4] Univ Strasbourg 1, F-67084 Strasbourg, France
关键词
D O I
10.1074/jbc.273.43.28444
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The neural chondroitin sulfate (CS) proteoglycan (PG) DSD-1-PG was originally identified with the monoclonal antibody (mAb) 473HD. It promotes neurite outgrowth of hippocampal neurons when coated as a substrate in the presence of polycations. This effect is inhibited by mAb 473HD that specifically recognizes the DSD-1 epitope. The DSD-1 epitope is also detectable in CS-C and CS-D preparations from shark cartilage but not in other chondroitin sulfates that are structurally related and differ in their sulfation patterns. Non-sulfated DSD-1-PG and chemically desulfated CS-D were not recognized by mAb 473HD, suggesting that the DSD-1 epitope depends on sulfation. It was possible to enrich DSD-1 epitope-bearing carbohydrates and D disaccharide units from CS-C and CS-D preparations on a mAb 473HD affinity matrix. This indicates that the DSD-1 epitope represents a distinct glycosaminoglycan structure containing D units. The analysis of glycosaminoglycan digestion products by high pressure liquid chromatography revealed that DSD-1-PG preparations contain a unique D disaccharide unit as well as an A, a C, and a non-sulfated disaccharide unit. In neurite outgrowth assays with hippocampal neurons, substrate-bound CS-D promoted neurite outgrowth, whereas CS-A, CS-B, or CS-C did not. This effect of CS-D was inhibited by mAb 473HD. DSD-1 epitope enriched fractions obtained from CS-D and CS-C promoted neurite outgrowth, whereas CS-C had no such effect prior to enrichment on the mAb 473HD matrix. Based on these findings we conclude that the DSD-1 epitope by itself is sufficient to promote neurite outgrowth and that this activity is possibly associated with D motifs.
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页码:28444 / 28453
页数:10
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