Journal
BIOPHYSICAL JOURNAL
Volume 117, Issue 1, Pages 46-55Publisher
CELL PRESS
DOI: 10.1016/j.bpj.2019.05.017
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Funding
- CERM/CIRMMP center of Instruct-ERIC, a Landmark European Strategy Forum on Research Infrastructures project
- Fondazione CR Firenze
- Italian Ministry of Foreign Affairs and International Cooperation [MAE0057283]
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Intrinsically disordered proteins (IDPs) as well as intrinsically disordered regions (IDRs) of complex protein machineries have recently been recognized as key players in many cellular functions. NMR represents a unique tool to access atomic resolution structural and dynamic information on highly flexible IDPs/IDRs. Improvements in instrumental sensitivity made heteronuclear direct detection possible for biomolecular NMR applications. The CON experiment has become one of the most useful NMR experiments to get a snapshot of an IDP/IDR in conditions approaching physiological ones. The availability of NMR spectrometers equipped with multiple receivers now enables the acquisition of several experiments simultaneously instead of one after the other. Here, we propose several variants of the CON experiment in which, during the recovery delay, a second two-dimensional experiment is acquired, either based on H-1 detection (CON//HN) or on N-15 detection (CON//btNH, CON//(H)CAN). The possibility to collect simultaneous snapshots of an IDP/IDR through different two-dimensional spectra provides a novel tool to follow chemical reactions, such as the occurrence of posttranslational modifications, as well as to study samples of limited lifetime such as cell lysates or whole cells.
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