Biophysical analysis of the interaction of laminin-related protein Netrin with its receptor Neogenin
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Date
2019-08
Authors
Sidhu, Shubleeen K
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Abstract
Netrins are axon guidance cue molecules belonging to the laminin related protein. These molecules play a significant role in neuronal migration, by releasing chemotrophic cues and allowing either chemoattraction or chemorepulsion. Netrin holds capability to bind multiple different receptors including Deleted in Colorectal Cancer (DCC), Uncoordinated 5 (UNC5) and Neogenin (Neo). In order to investigate the binding of Netrin with its receptor Neogenin, two different constructs of Neogenin, Neogenin 3-5 Long and Neogenin 3-5 Short were investigated with its binding partner Netrin in a biophysical approach followed by crystallization screening. Netrin and the two constructs of Neogenin alone and in complex were analysed by gel electrophoresis, Size exclusion chromatography, dynamic light scattering (DLS) and small angle X-ray scattering (SAXS). The findings from the low-resolution models reveal that the models for the complex appear larger and more compact than the individual models of the chNet△C, Neogenin Long 3-5 and Neogenin Short 3-5, which appear elongated. This analysis provides some insight into the interaction of Netrin with Neogenin and identifies 47% w/v 2-methyl-2,4- pentanediol 100mM HEPES, pH 7.5 and 35% w/v 2-methyl-2,4- pentanediol 100mM Imidazole, pH 8.0 as preliminary conditions for crystallization of the complex.
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Netrin