Proton clouds to measure long-range contacts between nonexchangeable side chain protons in solid-state NMR.
We show that selective labeling of proteins with protonated amino acids embedded in a perdeuterated matrix, dubbed 'proton clouds', provides general access to long-range contacts between nonexchangeable side chain protons in proton-detected solid-state NMR, which is important to study protein tertiary structure. Proton-cloud labeling significantly improves spectral resolution by simultaneously reducing proton line width and spectral crowding despite a high local proton density in clouds. The approach is amenable to almost all canonical amino acids. Our method is demonstrated on ubiquitin and the β-barrel membrane protein BamA.
NMR Spectroscopy, Bijvoet Center for Biomolecular Research, Department of Chemistry, Faculty of Science, Utrecht University , 3584 CH Utrecht, The Netherlands.
Tessa
T
Sinnige
Mark
M
Daniëls
Marc
M
Baldus
Markus
M
Weingarth
D000596:Amino Acids / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D011487:Protein Conformation; D011522:Protons; D025801:Ubiquitin / Q000737:chemistry*
D000596:Amino Acids; D011522:Protons; D025801:Ubiquitin