Investigation of dynamic solution interactions between NET-1 and UNC-5B by multi-wavelength analytical ultracentrifugation. 2023

Haben Gabir, and Monika Gupta, and Markus Meier, and Fabian Heide, and Manuel Koch, and Joerg Stetefeld, and Borries Demeler
Department of Chemistry, University of Manitoba, Winnipeg, MB, Canada.

NET-1 is a key chemotropic ligand that signals commissural axon migration and change in direction. NET-1 and its receptor UNC-5B switch axon growth cones from attraction to repulsion. The biophysical properties of the NET-1 + UNC-5B complex have been poorly characterized. Using multi-wavelength-AUC by adding a fluorophore to UNC-5B, we were able to separate the UNC-5B sedimentation from NET-1. Using both multi-wavelength- and single-wavelength AUC, we investigated NET-1 and UNC-5B hydrodynamic parameters and complex formation. The sedimentation velocity experiments show that NET-1 exists in a monomer-dimer equilibrium. A close study of the association shows that NET-1 forms a pH-sensitive dimer that interacts in an anti-parallel orientation. UNC-5B can form equimolar NET-1 + UNC-5B heterocomplexes with both monomeric and dimeric NET-1.

UI MeSH Term Description Entries
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000075386 Netrin Receptors Cell surface receptors that bind NETRINS. They typically contain both IMMUNOGLOBULIN DOMAINS and FIBRONECTIN TYPE III DOMAINS and function to mediate CELL MIGRATION and AXON GUIDANCE. Netrin Receptor,Netrin-1 Receptors,Receptor, Netrin,Netrin 1 Receptors,Receptors, Netrin,Receptors, Netrin-1
D000075388 Netrin-1 A netrin that binds the DCC RECEPTOR or UNC5 receptors, resulting respectively in axon attraction or repulsion. Its interaction with these receptors also prevents APOPTOSIS; it may function as a tumor suppressor protein. NTN1 Protein,Netrin-1 Protein,Netrin 1,Netrin 1 Protein,Protein, NTN1,Protein, Netrin-1
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D014461 Ultracentrifugation Centrifugation with a centrifuge that develops centrifugal fields of more than 100,000 times gravity. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
D054730 Protein Interaction Domains and Motifs Protein modules with conserved ligand-binding surfaces which mediate specific interaction functions in SIGNAL TRANSDUCTION PATHWAYS and the specific BINDING SITES of their cognate protein LIGANDS. Protein Interaction Domains,Protein Interaction Motifs,Binding Motifs, Protein Interaction,Protein Interaction Binding Motifs,Protein-Protein Interaction Domains,Domain, Protein Interaction,Domain, Protein-Protein Interaction,Domains, Protein Interaction,Domains, Protein-Protein Interaction,Motif, Protein Interaction,Motifs, Protein Interaction,Protein Interaction Domain,Protein Interaction Motif,Protein Protein Interaction Domains,Protein-Protein Interaction Domain

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