Increasing the sensitivity of cryoprobe protein NMR experiments by using the sole low-conductivity arginine glutamate salt.
Decrease in experimental sensitivity of cryoprobe experiments for salty samples, attributed to increased sample conductivity, has been a long-standing issue in protein NMR. Salt concentration can not be simply reduced as this often leads to protein aggregation. A simple and inexpensive solution to this problem is demonstrated here. We show that even for proteins prone to aggregation, the traditional solubilizing salt, 100mM NaCl, can be completely replaced by 50mM l-Arg and l-Glu. This replacement simultaneously reduces the sample conductivity and improves protein solubility. Up to a 6-fold overall increase in experimental sensitivity was achieved, in comparison with the traditional salty buffer. At constant protein concentration up to 2-fold increase in sensitivity was observed. The lengths of the proton pi/2 pulses were also significantly decreased, up to the level typical for non-salty samples in water.
Hautbergue
Guillaume M
GM
Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, UK.
Golovanov
Alexander P
AP
D003080:Cold Temperature; D004151:Dipeptides / Q000737:chemistry*; D004553:Electric Conductivity; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008832:Microchemistry / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004151:Dipeptides; D011506:Proteins; C024362:arginine glutamate