The effect of relaxation on the epitope mapping by saturation transfer difference NMR.
The effect of longitudinal relaxation of ligand protons on saturation transfer difference (STD) was investigated by using a known binding system, dihydrofolate reductase and trimethoprim. The results indicate that T1 relaxation of ligand protons has a severe interference on the epitope map derived from a STD measurement. When the T1s of individual ligand protons are distinctly different, STD experiments may not give an accurate epitope map for the ligand-target interactions. Measuring the relaxation times prior to mapping is strongly advised. A saturation time shorter than T1s is suggested for improving the potential epitope map. Reduction in temperature was seen to enhance the saturation efficiency in small to medium size targets.
Yan
Jiangli
J
Discovery Chemistry Research and Technologies, Lilly Research Labs, Lilly Corporate Center, Eli Lilly & Co, Indianapolis, IN 46285, USA.
Kline
Allen D
AD
Mo
Huaping
H
Shapiro
Michael J
MJ
Zartler
Edward R
ER
D000818:Animals; D002417:Cattle; D018604:Epitope Mapping / Q000379:methods*; D008099:Liver / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D011485:Protein Binding; D011522:Protons; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry*; D014295:Trimethoprim / Q000737:chemistry*
D011522:Protons; D014295:Trimethoprim; D013762:Tetrahydrofolate Dehydrogenase