Combined H-N Cross-Polarization and Carbonyl Detection NMR Spectroscopy Allow to Record High-Resolution, High-Sensitivity Spectra of Alpha-Synuclein in Bacterial Cells

Lopez J.M. · Methods in Molecular Biology (2023) · DOI: 10.1007/978-1-0716-2597-2_28

Resumen

Studies of intrinsically disordered proteins (IDPs) under physiological conditions by conventional NMR methods based on proton detection are severely limited by fast proton amide solvent exchange. Carbon detection has been proposed as a solution to the exchange problem but is hampered by low sensitivity. Here, we present a protocol combining proton–nitrogen cross-polarization and carbonyl detection to record high-resolution and high-sensitivity NMR spectra of IDPs under physiological conditions. The protocol describes a step-by-step method to register high-quality N-CO correlation spectrum of alpha-synuclein in E.coli bacterial cells at 37 °C.

Instituciones: Pontificia Universidad Católica del Perú