Epitope mapping of ligand-receptor interactions by diffusion NMR.
A novel method based on diffusion NMR for the epitope mapping of ligand binding is presented. The intermolecular NOE builds up during a long diffusion period and creates a deviation from the linearity. The ligand proton nearest the protein generates the strongest NOE from protein during the diffusion period and has the largest deviation. Therefore, this diffusion artifact can be used to characterize the ligand binding epitope. The concept was investigated using dihydrofolate reductase (DHFR) and its ligand trimethoprim (TMP), and the epitope map of TMP on DHFR generated with this method is in excellent agreement with the structural and dynamic studies by crystallography and NMR, as well as the medicinal chemistry results.
Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, Indiana 46285, USA.
Jiangli
J
Yan
Allen D
AD
Kline
Huaping
H
Mo
Edward R
ER
Zartler
Michael J
MJ
Shapiro
D018604:Epitope Mapping / Q000379:methods*; D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D012709:Serum Albumin / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D013762:Tetrahydrofolate Dehydrogenase / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D014295:Trimethoprim / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D014364:Tryptophan / Q000032:analysis / Q000276:immunology / Q000378:metabolism
D012709:Serum Albumin; D014364:Tryptophan; D014295:Trimethoprim; D013762:Tetrahydrofolate Dehydrogenase