Optimized pathway selection in intraresidual triple-resonance experiments.
An optimized intraresidual pulse sequence element with better sensitivity and suppression of sequential cross peaks is presented. Concatenation of three magnetization transfer delays allows their independent setting, in accordance with the relaxation properties of the individual spins, without concomitantly prolonging the pulse sequence. Additionally, implementations of the scheme to HNCA, HNCACB, and the TROSY based triple-resonance experiments are proposed. The feasibility of the new element was verified by recording HNCA and HNCACB on the small 8.6 kDa protein ubiquitin. The corresponding HNCA-TROSY experiment was tested on a larger protein, the 30.4 kDa Cel6A from the thermophilic soil bacterium Thermobifida fusca at 800 (1)H MHz.
Tossavainen
Helena
H
NMR Laboratory, Structural Biology and Biophysics Program, Institute of Biotechnology, University of Helsinki, P.O. Box 65, FIN-00014, Helsinki, Finland.
Permi
Perttu
P
D002247:Carbon Isotopes; D006859:Hydrogen; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D012815:Signal Processing, Computer-Assisted; D025801:Ubiquitin / Q000737:chemistry*
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D011506:Proteins; D025801:Ubiquitin; D006859:Hydrogen