Intrinsically disordered proteins (IDPs) have emerged as a central focus in structural biology due to their conformational flexibility and dynamic nature. Unlike classical proteins, IDPs do not adopt a single stable three-dimensional structure; instead, they exist as ensembles of rapidly interconverting conformations. This conformational malleability is crucial for their function and underlies their involvement in diverse cellular pathways. IDPs are ubiquitous in living organisms, and their malfunction is linked to the onset of severe diseases.

Researchers at CERM/CIRMMP have long been active in characterizing IDPs, exploiting different NMR techniques: a full array of experiments are now available to perform complete sequence specific assignment, to determine key observables to achieve structural and dynamic information and to acquire simultaneous snapshots of IDPs through multiple-receiver experiments. These techniques allow IDPs to be studied under near-physiological conditions (pH, temperature, ionic strength), and help identify functional modules enabled by disorder.

Several biomedically relevant IDPs have been studied at CERM. These include viral proteins, flexible linkers in large proteins such as CREB-binding protein (CBP) and the androgen receptor (AR), and α-synuclein in various physiologically relevant conditions. Other studies reveal sequence determinants of structural features, such as aromatic-proline motifs in osteopontin, which stabilize its compact states.

CERM has also advanced ultra-high field NMR protocols, highlighting the increased resolution achievable compared to lower magnetic fields. Hardware innovations, such as the use of multiple receivers combined with complementary labeling schemes, have enabled the investigation of interactions between cancer-related IDPs and large heterodimeric complexes. Novel methods have been developed to address the complexity of intrinsically disordered multidomain proteins. Additionally, CERM researchers contribute to the Protein Ensemble Database (PED) project, supporting the collection and dissemination of IDP conformational ensembles. A geometrical framework has also been developed to evaluate how well experimental observations match the conformational ensembles, providing a deeper understanding of IDP behavior.

 Watch on YouTube: IDPbyNMR the Movie

Selected publications:

Felli, I. C.; Pierattelli, R. Intrinsically Disordered Proteins Studied by NMR Spectroscopy; 2015. https://doi.org/10.1007/978-3-319-20164-1.

Bermel, W.; Bertini, I.; Felli, I. C.; Lee, Y.-M.; Luchinat, C.; Pierattelli, R. Protonless NMR Experiments for Sequence-Specific Assignment of Backbone Nuclei in Unfolded Proteins. J. Am. Chem. Soc. 2006, 128 (12), 3918–3919. https://doi.org/10.1021/ja0582206.

Schiavina, M.; Murrali, M. G.; Pontoriero, L.; Sainati, V.; Kümmerle, R.; Bermel, W.; Pierattelli, R.; Felli, I. C. Taking Simultaneous Snapshots of Intrinsically Disordered Proteins in Action. Biophys. J. 2019, 46–55. https://doi.org/10.1016/j.bpj.2019.05.017.

M. Schiavina, L. Bracaglia, M. A. Rodella, R. Kümmerle, R. Konrat, I. C. Felli, R. Pierattelli, Nat. Protocols. 2024, 19, 406–440. https://doi.org/10.1038/s41596-023-00921-9

M. Schiavina, L. Bracaglia, T. Bolognesi, M. A. Rodella, G. Tagliaferro, A. S. Tino, R. Pierattelli, I. C. Felli, J. Magn. Reson. Open 2024, 18, 100143. https://doi.org/10.1016/j.jmro.2023.100143

S. Knödlstorfer, M. Schiavina, M. A. Rodella, K. Ledolter, R. Konrat, R. Pierattelli, I. C. Felli, J.Am.Chem.Soc. 2024, 146, 27983–27987
https://doi.org/10.1021/jacs.4c09176?urlappend=%3Fref%3DPDF&jav=VoR&rel=cite-as

H. Ghafouri, T. Lazar, et al. Nucleic Acids Res., 2024, 52 (D1), D536–D544. 5)

L. Fiorucci, M. Schiavina, I. C. Felli, R. Pierattelli, E. Ravera, J. Chem. Inf. Model., 2024, 64, 5392–5401. https://pubs.acs.org/doi/10.1021/acs.jcim.4c00582

Bracaglia L, Oliveti S, Felli IC, Pierattelli R. Decoding Order and Disorder in Proteins by NMR Spectroscopy. J Am Chem Soc. 2025 Apr 23;147(16):13146-13157. doi: 10.1021/jacs.4c14959. Epub 2025 Apr 13. PMID: 40223218; PMCID: PMC12022988.

15N optimal control pulses: an efficient approach to enhance heteronuclear-detected NMR experiments at high magnetic fields.
Schiavina M, Joseph D, Griesinger C, Felli IC, Pierattelli R.J Magn Reson. 2025 Oct 4;381:107972. doi: 10.1016/j.jmr.2025.107972. Online ahead of print.PMID: 41066915

Exploring the Role of Structural and Dynamic Complexity in SARS-CoV-2 Nucleocapsid Protein-Heparin Interactions by NMR.
Bolognesi T, Schiavina M, Ciabini C, Parafioriti M, Gardini C, Elli S, Guerrini M, Pierattelli R, Felli IC.J Mol Biol. 2025 Sep 11:169437. doi: 10.1016/j.jmb.2025.169437. Online ahead of print.PMID: 40945576

NMR insights on multidomain proteins: the case of the SARS-CoV-2 nucleoprotein.
Bolognesi T, Schiavina M, Felli IC, Pierattelli R.Prog Nucl Magn Reson Spectrosc. 2025 Aug-Oct;148-149:101577. doi: 10.1016/j.pnmrs.2025.101577. Epub 2025 Jul 7.PMID: 40912879

Evidence of α-Synuclein/Glucocerebrosidase Dual Targeting by Iminosugar Derivatives.
Tagliaferro G, Davighi MG, Clemente F, Turchi F, Schiavina M, Matassini C, Goti A, Morrone A, Pierattelli R, Cardona F, Felli IC.ACS Chem Neurosci. 2025 Apr 2;16(7):1251-1257. doi: 10.1021/acschemneuro.4c00618. Epub 2025 Mar 13.PMID: 40079830

Revealing the Potential of a Chimaera: a Peptide-Peptide Nucleic Acid Molecule Designed To Interact with the SARS-CoV-2 Nucleocapsid Protein.
Tino AS, Quagliata M, Schiavina M, Pacini L, Papini AM, Felli IC, Pierattelli R.Angew Chem Int Ed Engl. 2025 Mar 10;64(11):e202420134. doi: 10.1002/anie.202420134. Epub 2025 Feb 14.PMID: 39912211

15N-detected TROSY for 1H-15N heteronuclear correlation to study intrinsically disordered proteins: strategies to increase spectral quality.
Rodella MA, Schneider R, Kümmerle R, Felli IC, Pierattelli R.J Biomol NMR. 2025 Mar;79(1):15-24. doi: 10.1007/s10858-024-00453-8. Epub 2025 Jan 22.PMID: 39841395 Free PMC article.

Disentangling the Complexity in Protein Complexes Using Complementary Isotope-Labeling and Multiple-Receiver NMR Spectroscopy.
Knödlstorfer S, Schiavina M, Rodella MA, Ledolter K, Konrat R, Pierattelli R, Felli IC.J Am Chem Soc. 2024 Oct 16;146(41):27983-27987. doi: 10.1021/jacs.4c09176. Epub 2024 Oct 7.PMID: 39374115

Minimum information guidelines for experiments structurally characterizing intrinsically disordered protein regions.
Mészáros B, Hatos A, Palopoli N, Quaglia F, Salladini E, Van Roey K, Arthanari H, Dosztányi Z, Felli IC, Fischer PD, Hoch JC, Jeffries CM, Longhi S, Maiani E, Orchard S, Pancsa R, Papaleo E, Pierattelli R, Piovesan D, Pritisanac I, Tenorio L, Viennet T, Tompa P, Vranken W, Tosatto SCE, Davey NE.Nat Methods. 2023 Sep;20(9):1291-1303. doi: 10.1038/s41592-023-01915-x. Epub 2023 Jul 3.PMID: 37400558 Review.

The Role of Disordered Regions in Orchestrating the Properties of Multidomain Proteins: The SARS-CoV-2 Nucleocapsid Protein and Its Interaction with Enoxaparin.
Schiavina M, Pontoriero L, Tagliaferro G, Pierattelli R, Felli IC.Biomolecules. 2022 Sep 15;12(9):1302. doi: 10.3390/biom12091302.PMID: 36139141 Free PMC article.

NMR Reveals Specific Tracts within the Intrinsically Disordered Regions of the SARS-CoV-2 Nucleocapsid Protein Involved in RNA Encountering.
Pontoriero L, Schiavina M, Korn SM, Schlundt A, Pierattelli R, Felli IC.Biomolecules. 2022 Jul 2;12(7):929. doi: 10.3390/biom12070929.