Normal carbon acid referencing for protein amide hydrogen exchange.
Measurement of the exchange kinetics for amide hydrogens along the protein backbone continues to offer valuable insights into structural stability and conformational dynamics. Since such studies routinely compare samples that differ in solution conditions or mechanical handling, normalization of the relative exchange rates can present a potentially significant source of experimental uncertainty. The carbon acids 1,3-dimethylimidazolium cation and thiomethylacetonitrile exhibit base catalyzed exchange rates similar to those of the slowly exchanging amides, under conditions typical for protein studies. With 13C enrichment at the acidic carbon position to facilitate selective observation, such carbon acids offer practical internal calibration of exchange.
LeMaster
David M
DM
Wadsworth Center, New York State Department of Health and Department of Biomedical Sciences, School of Public Health, University at Albany - SUNY, Empire State Plaza, Albany, New York, 12201 USA.
Anderson
Janet S
JS
Hernández
Griselda
G
D000577:Amides / Q000737:chemistry*; D002138:Calibration; D002247:Carbon Isotopes; D041961:Deuterium Exchange Measurement / Q000592:standards*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods / Q000592:standards*; D011506:Proteins / Q000737:chemistry*; D012015:Reference Standards; D013844:Thiazoles / Q000592:standards
D000577:Amides; D002247:Carbon Isotopes; D011506:Proteins; D013844:Thiazoles