2024-03-29T14:59:48Zhttps://eprints.lib.hokudai.ac.jp/dspace-oai/requestoai:eprints.lib.hokudai.ac.jp:2115/473852022-11-17T02:08:08Zhdl_2115_20055hdl_2115_8527An NMR strategy for fragment-based ligand screening utilizing a paramagnetic lanthanide probeSaio, TomohideOgura, KenjiShimizu, KazumiYokochi, MasashiBurke, Terrence R., Jr.1000070011757Inagaki, Fuyuhikoopen accessLigand screeningFragment-based drug design (FBDD)Protein-ligand structureLanthanide-binding peptide tag (LBT)Paramagnetic relaxation enhancement (PRE)Pseudo-contact shift (PCS)464A nuclear magnetic resonance-based ligand screening strategy utilizing a paramagnetic lanthanide probe is presented. By fixing a paramagnetic lanthanide ion to a target protein, a pseudo-contact shift (PCS) and a paramagnetic relaxation enhancement (PRE) can be observed for both the target protein and its bound ligand. Based on PRE and PCS information, the bound ligand is then screened from the compound library and the structure of the ligand-protein complex is determined. PRE is an isotropic paramagnetic effect observed within 30Å from the lanthanide ion, and is utilized for the ligand screening in the present study. PCS is an anisotropic paramagnetic effect providing long-range (∼40Å) distance and angular information on the observed nuclei relative to the paramagnetic lanthanide ion, and utilized for the structure determination of the ligand-protein complex. Since a two-point anchored lanthanide-binding peptide tag is utilized for fixing the lanthanide ion to the target protein, this screening method can be generally applied to non-metal-binding proteins. The usefulness of this strategy was demonstrated in the case of the growth factor receptor-bound protein 2 (Grb2) Src homology 2 (SH2) domain and its low- and high-affinity ligands.Springer Netherlands2011-11engjournal articleVoRhttp://hdl.handle.net/2115/47385https://doi.org/10.1007/s10858-011-9566-50925-2738Journal of Biomolecular NMR513395408https://eprints.lib.hokudai.ac.jp/dspace/bitstream/2115/47385/1/JBN51-3_395-408.pdfapplication/pdf2.22 MB2011-11