Site-specific proximity ligation provides molecular insights into biologically relevant interfaces of protein-protein interaction
| Autor: | Sung In Lim, Goeun Shin |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular 0301 basic medicine Azides Saccharomyces cerevisiae Proteins Multiprotein complex Biophysics Computational biology Crystallography X-Ray Ligands Biochemistry Protein–protein interaction 03 medical and health sciences Residue (chemistry) 0302 clinical medicine Molecular recognition Heterotrimeric G protein Protein Interaction Mapping Moiety Protein Interaction Domains and Motifs Amino Acids Nuclear pore Molecular Biology chemistry.chemical_classification Binding Sites Chemistry Cell Biology Amino acid Nuclear Pore Complex Proteins Cross-Linking Reagents 030104 developmental biology Multiprotein Complexes 030220 oncology & carcinogenesis Mutation Protein Binding |
| Zdroj: | Biochemical and Biophysical Research Communications. 533:932-937 |
| ISSN: | 0006-291X |
| DOI: | 10.1016/j.bbrc.2020.09.097 |
| Popis: | Dynamic protein-protein interactions (PPIs) are fundamental to spatiotemporal control of protein functions in biological systems. Dissecting binding interfaces in aqueous solution (i.e., biological interfaces) is of great importance for identifying molecular determinants that contribute to the affinity and specificity of PPIs. Herein, we describe a biochemical method, termed site-specific proximity ligation (SPL), that enables the identification and reconstruction of native binding interfaces distinct from those present in crystal structures and models from computational prediction. SPL involves the strategic incorporation of an aryl azide-containing unnatural amino acid (AZF) into residues of interest in a particular protein that forms a multiprotein complex. Depending on the interfacial role of a targeted residue, a photo-inducible highly reactive incorporated AZF moiety may react with neighboring functional groups to covalently capture an otherwise non-covalent or weak interaction with a specific partner protein, thereby revealing the landscape of biological interfaces. Using a heterotrimeric nuclear pore protein as a model, we show that the biological interfaces of the complex mapped by SPL provide new insight into dynamic molecular recognition that is missed by, or even in conflict with, static models. |
| Databáze: | OpenAIRE |
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