Characterization of a sulfoglucuronyl carbohydrate binding protein in the developing nervous system.
Characterization of a sulfoglucuronyl carbohydrate binding protein in the developing nervous system.
复制标题
发育中的神经系统中磺基葡萄糖醛酸糖结合蛋白的表征。
DOI：
10.1046/j.1471-4159.1997.68031286.x
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发表时间：
1997
影响因子：
4.7
通讯作者：
Jungalwala,FB
中科院分区：
文献类型：
--
作者：
Nair,SM;Jungalwala,FB
在磺基葡萄糖醛酸糖脂（SGGL）和某些糖蛋白上发现的发育调节和阶段特异性表达的HNK-1碳水化合物已被提出通过其与蛋白质“受体”的相互作用参与神经细胞粘附和识别过程。我们从新生大鼠脑中分离并纯化了一种约30 kDa的SGGL结合蛋白（SBP-1）。SBP-1在固相免疫结合和高效薄层色谱免疫覆盖试验中均与SGGL和硫苷脂特异性结合。N-末端序列分析显示SBP-1类似于促进粘附性神经突生长的蛋白质Escherin。SGGL的脱硫导致SBP-1结合的消除。然而，通过酯化或还原羧基对葡萄糖醛酸部分进行化学修饰没有影响，表明SBP-1结合需要碳水化合物连接的硫酸酯基团。SBP-1与SGGL的结合被HNK-1抗体特异性抑制，但不被其他IgM抗体抑制。然而，SBP-1与硫苷脂的结合不被HNK-1抗体抑制。肝素、岩藻依聚糖和硫酸葡聚糖（50 K）也抑制SBP-1与SGGL的结合。在大鼠大脑皮层发育过程中，SBP-1的水平在胚胎第18天后下降到出生后第10天几乎检测不到的水平;而在小脑中，SBP-1的表达在出生后第7天达到最大。SBP-1还与通过HNK-1免疫亲和层析从大鼠脑中分离的HNK-1糖蛋白特异性结合。不含HNK-1碳水化合物的蛋白质不结合SBP-1。与HNK-1糖蛋白的结合被HNK-1抗体抑制，但不被其他IgM抗体抑制，表明结合是通过蛋白质的HNK-1碳水化合物部分介导的。SBP-1与SGGL和HNK-1糖蛋白在围产期脑发育期间的相互作用和共表达表明该蛋白具有功能性作用。
The developmentally regulated and stage‐specifically expressed HNK‐1 carbohydrate found on sulfoglucuronylglycolipids (SGGLs) and certain glycoproteins has been proposed to be involved in neural cell adhesion and recognition processes through its interaction with protein “receptors.” We have isolated and purified a ∼30‐kDa SGGL‐binding protein (SBP‐1) from neonatal rat brain. SBP‐1 specifically bound to SGGLs and sulfatide both in solid‐phase immunobinding and high‐performance thin‐layer chromatography‐immunooverlay assays. N‐terminal sequence analysis showed that SBP‐1 is similar to an adhesive neurite outgrowth promoting protein amphoterin. Desulfation of SGGLs resulted in abolition of SBP‐1 binding. However, chemical modification of glucuronic acid moiety by either esterification or reduction of the carboxyl group had no effect, suggesting requirement of the carbohydrate‐linked sulfate group for SBP‐1 binding. The binding of SBP‐1 to SGGLs was specifically inhibited by HNK‐1 antibody but not by other IgM antibodies. The binding of SBP‐1 to sulfatide, however, was not inhibited by HNK‐1 antibody. Heparin, fucoidan, and dextran sulfate (50K) also inhibited the binding of SBP‐1 to SGGLs. During development of the rat cerebral cortex, the level of SBP‐1 decreased after embryonic day 18 to an almost undetectable level by postnatal day 10; whereas in the cerebellum, the expression of SBP‐1 was maximal at postnatal day 7. SBP‐1 also bound specifically to the HNK‐1 glycoproteins isolated from rat brain by HNK‐1 immunoaffinity chromatography. Proteins without HNK‐1 carbohydrate did not bind SBP‐1. The binding to HNK‐1 glycoproteins was inhibited by HNK‐1 antibody, but not by other IgM antibodies, indicating that the binding was mediated through the HNK‐1 carbohydrate moiety of the proteins. The interaction and coexpression of SBP‐1 with SGGLs and HNK‐1 glycoproteins, during the perinatal brain development, suggest a functional role for this protein.