Hierarchical folding-upon-binding of an intrinsically disordered protein
Intrinsically disordered proteins (IDPs) often undergo folding-upon-binding to their partners via short linear motifs, typically 5-15 amino acids in length. However, a significant proportion of IDPs do not adhere to this paradigm but fold upon binding through extended regions comprising multiple molecular recognition elements. For these IDPs, the binding mechanisms and the structural characteristics of their folding intermediates remain poorly understood. Here we unveil hierarchical folding of an IDP as it binds to its partner, exemplified by the disordered signaling effector POSH and the small GTPase Rac1. By combining nuclear magnetic resonance (NMR) spectroscopy and X-ray crystallography, we resolve at atomic resolution how POSH transitions from a fully disordered state to a highly ordered, Rac1-bound conformation through two structurally distinct folding intermediates. The folding of each element is contingent on the successful structuring of the preceding element, highlighting a hierarchical folding-upon-binding mechanism. Our work highlights the potential of targeting folding intermediates and conformational transitions to unlock therapeutic opportunities for IDPs.
Authors
- Andrés Palencia (ORCID: https://orcid.org/0000-0002-1805-319X)
- Malene Ringkjøbing Jensen (ORCID: https://orcid.org/0000-0003-0419-2196)
- Elisabetta Boeri Erba (ORCID: https://orcid.org/0000-0002-5910-4707)
- Elise Delaforge (ORCID: https://orcid.org/0000-0001-8014-5100)
- Lenette F. Kjaer (ORCID: https://orcid.org/0000-0002-0847-4169)
- Maud Tengo
- Guillaume Bouvignies (ORCID: https://orcid.org/0000-0003-4398-0320)
- Francesco Ielasi (ORCID: https://orcid.org/0000-0002-5272-0865)
- Luiza M. Bessa (ORCID: https://orcid.org/0000-0002-1993-3105)
- Laura Mariño (ORCID: https://orcid.org/0000-0003-1658-7179)
- Thomas Winbolt (ORCID: https://orcid.org/0009-0009-7670-2336)
Institutions
- Centre National de la Recherche Scientifique (FR)
- Chimie ParisTech - PSL (FR)
- Inserm (FR)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
- Université Paris Sciences et Lettres (FR)
- École Normale Supérieure - PSL (FR)
- CEA Grenoble (FR)
- Sorbonne Université (FR)
- Servei de Salut de les Illes Balears (ES)
- Departament de Salut (ES)
- Grenoble Institute of Neurosciences (FR)
- Institut de Biologie Structurale (FR)
- Institut pour l'avancée des biosciences (FR)
- Université Grenoble Alpes (FR)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-11-27
- DOI
- https://doi.org/10.1038/s41467-025-66420-5
- Citations
- 3
- Primary Topic
- Protein Structure and Dynamics
- Type
- article
- Field-Weighted Citation Impact
- 2.42