Glycosaminoglycan-binding properties and secondary structure of the C-terminus of netrin-1
Glycosaminoglycan-binding properties and secondary structure of the C-terminus of netrin-1
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DOI：
10.1006/bbrc.2000.2583
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发表时间：
2000-05-10
影响因子：
3.1
通讯作者：
Koch, KW
中科院分区：
文献类型：
--
作者：
Kappler, J;Franken, S;Koch, KW
Netrins是一种与层粘连蛋白B2链相关的可溶性轴突生长促进蛋白。由于这些蛋白质及其受体DCC(在结直肠癌中缺失的基因产物)与肝素结合，糖胺多糖可能会以类似于其他几种配体-受体系统的方式调节它们的生物学作用。根据圆二色谱实验，我们证明了含有Netrin-1的C-末端碱性氨基酸簇的多肽(I)在水-三氟乙醇混合物中采用α-螺旋构象，(II)在生理离子条件下以高亲和力与肝素静电结合(根据表面等离子体共振K(D)=15 nM与固定化肝素结合，用等温滴定量热法测定K(D)=50 nM)。这些数据表明，Netrin-1 C末端的碱性氨基酸簇形成了一个α-螺旋结构元件，它可能有助于这种神经营养导向分子的糖胺聚糖结合活性。(C)2000年学术出版社。
Netrins are soluble neurite-outgrowth-promoting proteins related to the laminin B2 chain. Since these proteins and their receptor DCC (the "deleted in colorectal carcinoma" gene product) bind heparin, glycosaminoglycans may modulate their biological actions in a similar fashion as described for several other ligand-receptor systems. Here we show that a polypeptide encompassing the C-terminal cluster of basic amino acids of netrin-1 (i) adopts an alpha-helical conformation in water-trifluoroethanol mixtures according to circular dichroism experiments and (ii) binds electrostatically to heparin with high affinity under physiological ionic conditions (K(D) = 15 nM for the binding to immobilized heparin according to surface plasmon resonance, K(D) = 50 nM in solution as determined with isothermal titration calorimetry). These data indicate that the cluster of basic amino acids at the C-terminus of netrin-1 forms an alpha-helical structural element which can contribute to the glycosaminoglycan-binding activity of this neurotrophic guidance molecule. (C) 2000 Academic Press.