Isolation of a Tenascin-R Binding Protein from Mouse Brain Membranes
Isolation of a Tenascin-R Binding Protein from Mouse Brain Membranes
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DOI：
10.1074/jbc.272.51.32092
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发表时间：
1997-12
期刊：
影响因子：
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通讯作者：
Zhi‐cheng Xiao;U. Bartsch;R. Margolis;G. Rougon;D. Montag;M. Schachner
中科院分区：
文献类型：
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作者：
Zhi‐cheng Xiao;U. Bartsch;R. Margolis;G. Rougon;D. Montag;M. Schachner
我们已经分离出硫酸软骨素蛋白聚糖从小鼠脑细胞外基质糖蛋白腱生蛋白-R（TN-R），包括氨基末端富含半胱氨酸的拉伸和4.5表皮生长因子样重复片段的亲和层析。分离的硫酸软骨素蛋白聚糖具有500-600 kDa的分子量，并携带HNK-1碳水化合物表位。用软骨素酶ABC处理显示约400 kDa的主带和200和150 kDa的两个次带。免疫印迹分析涉及磷酸蛋白聚糖的分子，但不是硫酸软骨素蛋白聚糖，神经聚糖和多功能蛋白聚糖。磷酸根聚糖相关分子与TN-R的表皮生长因子样重复序列的结合是钙依赖性的。通过免疫细胞化学，该分子与TN-R在视网膜和视神经中的共定位表明这两种分子在体内的功能关系。磷酸根聚糖相关分子在体外对海马神经元轴突生长的抑制作用在作为均匀基质涂覆时被TN-R中和。此外，磷酸根聚糖相关分子中和由TN-R包被的尖锐底物边界诱导的生长锥排斥，有或没有预先用软骨素酶ABC处理。这些观察结果表明，TN-R可以相互作用与磷酸根相关的分子，从而调节其对轴突发生的抑制作用。
We have isolated a chondroitin sulfate proteoglycan from mouse brain by affinity chromatography with a fragment of the extracellular matrix glycoprotein tenascin-R (TN-R) that comprises the amino-terminal cysteine-rich stretch and the 4.5 epidermal growth factor-like repeats. The isolated chondroitin sulfate proteoglycan has a molecular mass of 500–600 kDa and carries the HNK-1 carbohydrate epitope. Treatment with chondroitinase ABC reveals a major band of approximately 400 kDa and two minor bands at 200 and 150 kDa. Immunoblot analysis relates the molecule to phosphacan but not to the chondroitin sulfate proteoglycans neurocan and versican. Binding of the phosphacan-related molecule to the epidermal growth factor-like repeats of TN-R is Ca2+-dependent. Co-localization of the molecule with TN-R in the retina and optic nerve by immunocytochemistry suggests a functional relationship between the two molecules in vivo. Inhibition of neurite outgrowth from hippocampal neurons by the phosphacan-related molecule in vitro is neutralized by TN-R when coated as a uniform substrate. Furthermore, the phosphacan-related molecule neutralizes growth cone repulsion induced by TN-R coated as a sharp substrate boundary with or without prior treatment with chondroitinase ABC. These observations indicate that TN-R can interact with a phosphacan-related molecule and thereby modulate its inhibitory influence on neuritogenesis.