caa a22 4500 445325119 CHVBK 20180317142717.0 cr unu---uuuuu 170323e20111101xx s 000 0 eng 10.1007/s10858-011-9566-5 doi (NATIONALLICENCE)springer-10.1007/s10858-011-9566-5 An NMR strategy for fragment-based ligand screening utilizing a paramagnetic lanthanide probe [Elektronische Daten] [Tomohide Saio, Kenji Ogura, Kazumi Shimizu, Masashi Yokochi, Terrence Burke Jr., Fuyuhiko Inagaki] A 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. The Author(s), 2011 Ligand screening nationallicence Fragment-based drug design (FBDD) nationallicence Protein-ligand structure nationallicence Lanthanide-binding peptide tag (LBT) nationallicence Paramagnetic relaxation enhancement (PRE) nationallicence Pseudo-contact shift (PCS) nationallicence Saio Tomohide Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut Ogura Kenji Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut Shimizu Kazumi Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut Yokochi Masashi Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut Burke Jr. Terrence National Cancer Institute at Frederick, Laboratory of Medicinal Chemistry, Center for Cancer Research, P. O. Box B, 21702-1201, Frederick, MD, USA aut Inagaki Fuyuhiko Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 395-408 0925-2738 51:3<395 2011 51 10858 https://doi.org/10.1007/s10858-011-9566-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9566-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Saio Tomohide Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut NATIONALLICENCE 700 1- Ogura Kenji Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut NATIONALLICENCE 700 1- Shimizu Kazumi Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut NATIONALLICENCE 700 1- Yokochi Masashi Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut NATIONALLICENCE 700 1- Burke Jr Terrence National Cancer Institute at Frederick, Laboratory of Medicinal Chemistry, Center for Cancer Research, P. O. Box B, 21702-1201, Frederick, MD, USA aut NATIONALLICENCE 700 1- Inagaki Fuyuhiko Department of Structural Biology, Faculty of Advanced Life Science, Hokkaido University, N-21, W-11, Kita-ku, 001-0021, Sapporo, Japan aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 51/3(2011-11-01), 395-408 0925-2738 51:3<395 2011 51 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer