Interplay between partner and ligand facilitates the folding and binding of an intrinsically disordered protein
| Autor: | Christopher M. Baker, Chi T. Wong, David De Sancho, Joseph M. Rogers, Sarah L. Shammas, Jane Clarke, Vladimiras Oleinikovas |
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| Přispěvatelé: | Clarke, Jane [0000-0002-7921-900X], Apollo - University of Cambridge Repository |
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Protein Folding
BCL-2 protein–protein interactions Biology Molecular Dynamics Simulation Intrinsically disordered proteins Ligands coarse-grained simulation Protein Structure Secondary Protein–protein interaction stopped flow Protein structure Proto-Oncogene Proteins Multidisciplinary Binding protein Protein engineering Biological Sciences Protein tertiary structure Folding (chemistry) Intrinsically Disordered Proteins Crystallography Kinetics Mutation Biophysics Myeloid Cell Leukemia Sequence 1 Protein Protein folding Mutant Proteins Protein Binding |
| Popis: | Protein–protein interactions are at the heart of regulatory and signaling processes in the cell. In many interactions, one or both proteins are disordered before association. However, this disorder in the unbound state does not prevent many of these proteins folding to a well-defined, ordered structure in the bound state. Here we examine a typical system, where a small disordered protein (PUMA, p53 upregulated modulator of apoptosis) folds to an α-helix when bound to a groove on the surface of a folded protein (MCL-1, induced myeloid leukemia cell differentiation protein). We follow the association of these proteins using rapid-mixing stopped flow, and examine how the kinetic behavior is perturbed by denaturant and carefully chosen mutations. We demonstrate the utility of methods developed for the study of monomeric protein folding, including β-Tanford values, Leffler α, Φ-value analysis, and coarse-grained simulations, and propose a self-consistent mechanism for binding. Folding of the disordered protein before binding does not appear to be required and few, if any, specific interactions are required to commit to association. The majority of PUMA folding occurs after the transition state, in the presence of MCL-1. We also examine the role of the side chains of folded MCL-1 that make up the binding groove and find that many favor equilibrium binding but, surprisingly, inhibit the association process. |
| Databáze: | OpenAIRE |
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