| 초록 |
In biological systems, protein-protein interactions (PPIs) are important for spatiotemporal regulation of protein functions. To identify molecular contribution to specific interactions in a native condition, it is important to dissect PPI interfaces in solution phase (i.e., biological interfaces). Here we introduce a biochemical approach which makes it possible to recognize native binding interfaces distinct from those present in crystal structures and computer-predicted models. The strategy incorporates p-azido-phenylalanine (AZF) into a residue of interest in a particular protein that forms a multiproteins complex. Depending on the role of targeted residue, photo-inducible AZF may covalently bind with partner's residues. Using a heterotrimeric nuclear pore protein complex as a model, we demonstrate a new landscape of the biological interfaces of the complex, offering new perspective into dynamic molecular recognition that is missed by crystal structure and static models. |
