Understanding Protein Function Through an Ensemble Description: Characterization of Functional States by 19F NMR

Perfluorocarbons in Chemical Biology

“…Many have used the above protein fluorination methods to monitor protein dynamics and proteinmembrane interactions by NMR. [67][68][69]79,80] Mehl, Pielak, and coworkers have used site-specific incorporation of trifluoromethylphenylalanine 7to study protein conformation and binding, [50] as well as detect proteins in E. coli. [51] Whereas monofluorinated AAs resulted in broadened fluorine signal that was unable to be detected in large proteins, the fluorine signal from 7 could be detected even for proteins up to 100 kDa, demonstrating the need to incorporate perfluorinated groups.…”

“…The addition of multiple, chemically equivalent fluorine atoms (i. e., trifluoromethyl, hexafluoroisopropyl, or perfluoro‐ tert‐ butyl) enhances the sensitivity of 19 F protein NMR experiments. Many have used the above protein fluorination methods to monitor protein dynamics and protein‐membrane interactions by NMR [67–69,79,80] . Mehl, Pielak, and co‐workers have used site‐specific incorporation of trifluoromethylphenylalanine ( 7 ) to study protein conformation and binding, [50] as well as detect proteins in E. coli [51] .…”

Predicting 19F NMR Chemical Shifts: A Combined Computational and Experimental Study of a Trypanosomal Oxidoreductase–Inhibitor Complex

“…In combination with the 200 sampling points taken from each of the individual MD trajectories, this makes it one of the largest QM-based 19 F NMR chemical-shift calculations reported so far. [7,8,27] The QM size and the combined number of calculations in this context are unprecedented and are at the frontier of what is currently possible for such large biological systems. The embedded QM regions are shown in Figure 1 C-F. A detailed description is given in the Supporting Information (Supporting Information, Table S3).…”

Monitoring Dynamics of Large Membrane Proteins by 19F Paramagnetic Longitudinal Relaxation: Domain Movement in a Glutamate Transporter Homolog

“…19 F-NMR is a useful alternative because of its high sensitivity, robust chemical shift dispersion, the absence of background signals, and no requirement for perdeuteration 5 6 . Fluorinated probes have been introduced into proteins via incorporation of fluorinated or unnatural amino acids 7 or via cysteine chemistry 8 . Many methods to follow dynamics, including saturation transfer, EXSY, CPMG, CEST and DEST, have been adapted to 19 F-NMR 6,[9][10][11][12] .…”

“…Typically, the assignment relies on population shifts expected in response to ligand binding or mutations and, therefore, requires prior information from other techniques 9 . Alternatively, differences in solvent paramagnetic resonance enhancement (PRE) 8 have been used, but this approach only works when the conformational states feature distinct solvent exposure of the 19 F probe. Here, we undertook measurements of 19 F longitudinal relaxation rates (R1) and their distancedependent enhancement by paramagnetic ions chelated by di-histidine motifs to assign resonances to different states and to estimate the rates of the conformational exchange.…”