A novel approach for studying receptor-ligand interactions on living cells surface by using NUS/T1ρ-NMR methodologies combined with computational techniques: The RGDechi15D-α v β 5 integrin complex.

Autor: Farina B; Institute of Biostructures and Bioimaging-CNR, Via Mezzocannone 16, 80134 Naples, Italy.; Advanced Accelerator Applications, a Novartis Company, via Vivaldi 43, 81100 Caserta, Italy., Andrea C; Department of Environmental, Biological and Pharmaceutical Science and Technology, University of Campania - Luigi Vanvitelli, via Vivaldi 43, 81100 Caserta, Italy., Del Gatto A; Institute of Biostructures and Bioimaging-CNR, Via Mezzocannone 16, 80134 Naples, Italy.; Interdepartmental Center of Bioactive Peptide, University of Naples Federico II, Via Mezzocannone 16, 80134 Naples, Italy., Comegna D; Institute of Biostructures and Bioimaging-CNR, Via Mezzocannone 16, 80134 Naples, Italy., Di Gaetano S; Institute of Biostructures and Bioimaging-CNR, Via Mezzocannone 16, 80134 Naples, Italy.; Interdepartmental Center of Bioactive Peptide, University of Naples Federico II, Via Mezzocannone 16, 80134 Naples, Italy., Capasso D; Interdepartmental Center of Bioactive Peptide, University of Naples Federico II, Via Mezzocannone 16, 80134 Naples, Italy.; Center for Life Sciences and Technologies (CESTEV) University of Naples Federico II, Via Tommaso De Amicis 95, 80145 Naples, Italy., Paladino A; Department of Science and Technology, University of Sannio, via Francesco de Sanctis, Benevento 82100, Italy., Acconcia C; Department of Environmental, Biological and Pharmaceutical Science and Technology, University of Campania - Luigi Vanvitelli, via Vivaldi 43, 81100 Caserta, Italy., Gentile MT; Department of Environmental, Biological and Pharmaceutical Science and Technology, University of Campania - Luigi Vanvitelli, via Vivaldi 43, 81100 Caserta, Italy., Saviano M; Institute of Crystallography-CNR, Via Amendola 122/O, 70126 Bari, Italy., Fattorusso R; Department of Environmental, Biological and Pharmaceutical Science and Technology, University of Campania - Luigi Vanvitelli, via Vivaldi 43, 81100 Caserta, Italy.; Interdepartmental Center of Bioactive Peptide, University of Naples Federico II, Via Mezzocannone 16, 80134 Naples, Italy., Zaccaro L; Institute of Biostructures and Bioimaging-CNR, Via Mezzocannone 16, 80134 Naples, Italy.; Interdepartmental Center of Bioactive Peptide, University of Naples Federico II, Via Mezzocannone 16, 80134 Naples, Italy., Russo L; Department of Environmental, Biological and Pharmaceutical Science and Technology, University of Campania - Luigi Vanvitelli, via Vivaldi 43, 81100 Caserta, Italy.
Jazyk: angličtina
Zdroj: Computational and structural biotechnology journal [Comput Struct Biotechnol J] 2021 May 29; Vol. 19, pp. 3303-3318. Date of Electronic Publication: 2021 May 29 (Print Publication: 2021).
DOI: 10.1016/j.csbj.2021.05.047
Abstrakt: Structural investigations of receptor-ligand interactions on living cells surface by high-resolution Nuclear Magnetic Resonance (NMR) are problematic due to their short lifetime, which often prevents the acquisition of experiments longer than few hours. To overcome these limitations, we developed an on-cell NMR-based approach for exploring the molecular determinants driving the receptor-ligand recognition mechanism under native conditions. Our method relies on the combination of high-resolution structural and dynamics NMR data with Molecular Dynamics simulations and Molecular Docking studies. The key point of our strategy is the use of Non Uniform Sampling (NUS) and T1ρ-NMR techniques to collect atomic-resolution structural and dynamics information on the receptor-ligand interactions with living cells, that can be used as conformational constraints in computational studies. In fact, the application of these two NMR methodologies allows to record spectra with high S/N ratio and resolution within the lifetime of cells. In particular, 2D NUS [ 1 H- 1 H] trNOESY spectra are used to explore the ligand conformational changes induced by receptor binding; whereas T1ρ-based experiments are applied to characterize the ligand binding epitope by defining two parameters: T1ρ Attenuation factor and T1ρ Binding Effect. This approach has been tested to characterize the molecular determinants regulating the recognition mechanism of α v β 5 -integrin by a selective cyclic binder peptide named RGDechi15D. Our data demonstrate that the developed strategy represents an alternative in-cell NMR tool for studying, at atomic resolution, receptor-ligand recognition mechanism on living cells surface. Additionally, our application may be extremely useful for screening of the interaction profiling of drugs with their therapeutic targets in their native cellular environment.
Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(© 2021 The Author(s).)
Databáze: MEDLINE
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