H2020Индивидуална стипендия2019–2021

evoDISFOLD · Decoding Coupled Folding and Binding via Single-Molecule characterization of Ancient IDP protein-protein Interactions

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2019-09-01 → 2021-08-31
Финансиране от ЕС: 175 420 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Безоформените протеини, като NCBD, се променят и придобиват конкретна структура, когато се свързват с други молекули. Разбирането на този процес помага да се разбере как работят сигнализацията и регулацията в клетките.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

Decoding Coupled Folding and Binding via Single-Molecule characterization of Ancient IDP protein-protein Interactions

Project Context: Intrinsically Disordered Proteins (IDPs) lack a defined 3-dimensional structure under physiological conditions, and can perform their functions as disordered ensembles. They are now acknowledged as extremely relevant players of cellular processes because of their abundance in proteomes, and also because they are associated with crucial functions of signaling and regulation. The historical neglect of IDPs is in part explained by the limitations of classical biophysical techniques for resolving IDPs conformational heterogeneity and dynamics. Nonetheless, in the last decade single molecule techniques like fluorescence spectroscopy using Förster Resonance Energy Transfer (smFRET) have greatly advanced the understanding and modelling of IDPs complexity. Some IDPs develop their activity as fully disordered ensembles, but many also undergo different degrees of disorder-to-order transition when they bind other molecules to perform their function—this process is known as coupled folding and binding. A widely studied example is that of the nuclear co-activator binding domain of transcriptional co-activators CREB-binding protein and p300 (NCBD), upon interaction with its multiple binding partners. Overall Objectives: Our group pioneered the study of IDPs using smFRET—in particular, it has extensively studied the coupled folding and binding of NCBD with several of its partners, some also intrinsically disordered like the CREBBP interacting domain of Nuclear Receptor Coactivators 3 (ACTR) and 1 (SCR-1), or the transactivation domain of p53 (p53TAD). The overall objective of the project was to expand the possibilities to study IDPs with single-molecule techniques by developing methods and protocols allowing to use Optical Tweezers force spectroscopy to analyze the coupled folding and binding of NCBD with its interaction partners. With this achievement, we sought to obtain new orthogonal information and to progress towards the integration of high resolution force and smFRET data—with the ultimate goal of disentangling general principles of coupled folding and binding.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

Intrinsically Disordered Proteins (IDPs) are characterized by their inability to spontaneously fold into a defined 3D structure. Nonetheless, many IDPs are able to undergo disorder-to-order transitions upon binding to interaction partners, in a process known as coupled folding and binding. Although different mechanisms have attempted to rationalize this process, technical limitations have prevented from deciphering the underlying principles of coupled folding and binding.Here, I will tackle this biological problem by introducing a novel, highly interdisciplinary approach integrating, for the first time, optical tweezers (OT) and single-molecule FRET (smFRET) to resolve the microscopic steps in coupled folding and binding; and Ancestral Sequence Reconstruction (ASR) as a rational strategy to guide protein engineering.For this purpose, I will study the nuclear co-activator binding domain (NCBD) and its multiple polypeptide binding partners (both intrinsically disordered and structured) as a model to resurrect, using customized ASR methods, the sequences of interacting pairs at multiple divergence nodes along a wide evolutionary scale. Then, I will characterize their intermolecular binding and intramolecular conformational dynamics with OT/smFRET.This cutting-edge project will result in the determination of mutational trajectories defining i) the emergence of coupled folding and binding, ii) the evolution of this process, and iii) the impact of promiscuous binding to multiple partners on its evolution. This data will allow to extract the essential determinants of coupled folding and binding.

Оригинален текст от CORDIS (на английски).

Участници

UNIVERSITAT ZURICH · ZurichКоординаторШвейцария