Investigation of dynamic solution interactions between NET-1 and UNC-5B by multi-wavelength analytical ultracentrifugation
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Date
2023
Authors
Gabir, Haben
Gupta, Monika
Meier, Markus
Heide, Fabian
Koch, Manuel
Stetefeld, Joerg
Demeler, Borries
Journal Title
Journal ISSN
Volume Title
Publisher
Springer Link
Abstract
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.
Description
Accepted author manuscript
Keywords
Netrin-1 , Uncoordinated-5 , Hetero-interaction studies between proteins , Multi-wavelength analytical ultracentrifugation , UltraScan , UltraScan-SOMO , Hydrodynamic bead modeling , Axon guidance
Citation
Gabir, H., Gupta, M., Meier, M., Heide, F., Koch, M., Stetefeld, J., & Demeler, B. (2023). Investigation of dynamic solution interactions between NET-1 and UNC-5B by multi-wavelength analytical ultracentrifugation. European Biophysics Journal, 52(4-5), 473-481.https://doi.org/10.1007/s00249-023-01644-