Glycosaminoglycan-binding properties and secondary structure of the C-terminus of netrin-1

被引:57
作者
Kappler, J
Franken, S
Junghans, U
Hoffmann, R
Linke, T
Müller, HW
Koch, KW
机构
[1] Univ Bonn, Inst Physiol Chem, D-53115 Bonn, Germany
[2] Univ Dusseldorf, Neurol Klin, Mol Neurobiol Lab, D-40225 Dusseldorf, Germany
[3] Univ Dusseldorf, Biol Med Forschungszentrum, D-40225 Dusseldorf, Germany
[4] Forschungszentrum Julich, Inst Biol Informat Verarbeitung, D-52425 Julich, Germany
[5] Univ Bonn, Kekule Inst Organ & Biochem, D-58121 Bonn, Germany
关键词
proteoglycans; nervous system development; extracellular matrix;
D O I
10.1006/bbrc.2000.2583
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
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.
引用
收藏
页码:287 / 291
页数:5
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