Discovering High-Affinity Ligands for Proteins: SAR by NMR

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Science  29 Nov 1996:
Vol. 274, Issue 5292, pp. 1531-1534
DOI: 10.1126/science.274.5292.1531


  • Fig. 1.

    An outline of the SAR by NMR method.

  • Fig. 2.

    A superposition of 15N-HSQC spectra for FKBP in the absence (magenta contours) and presence (black contours) of compound 3. Both spectra were acquired in the presence of saturating amounts of 2 (2.0 mM). Significant chemical shifts changes are observed for labeled residues.

  • Fig. 3.

    A surface representation of FKBP showing the locations of 2 and 9, as determined from 15N-13C-filtered NOE data (11). Residues that exhibited the largest chemical shift changes on the binding of 2, 9, or both 2 and 9 are colored in magenta, cyan, and yellow, respectively. Chemical shift changes for 9 (cyan and yellow) are those observed on the addition of 9 to FKBP in the presence of saturating amounts of 2 (2.0 mM). Weighted averaged chemical shifts were used (Δδ(1H, 15N) = |Δδ(1H)| + 0.2*|Δδ(15N)|), and colored residues are those for which Δδ(1H, 15N) exceeded 0.15 and 0.05 ppm for 2 and 9, respectively.

  • Fig. 4.

    Summary of the SAR by NMR method as applied to FKBP. Compounds with affinities from 19 to 228 nM were obtained from two fragment leads with affinities of 2 and 100 μM.

  • Fig. 5.

    Ribbon (23) depiction of the structure of FKBP (gray) when complexed to 14 (green carbon atoms). Shown in yellow are those residues that have NOEs to the ligand.

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