2 research outputs found
Rapid Identification of Ligand-Binding Sites by Using an Assignment-Free NMR Approach
In this study, we developed an assignment-free
approach for rapid
identification of ligand-binding sites in target proteins by using
NMR. With a sophisticated cell-free stable isotope-labeling procedure
that introduces <sup>15</sup>N- or <sup>13</sup>C-labels to specific
atoms of target proteins, this approach requires only a single series
of ligand titrations with labeled targets. Using titration data, ligand-binding
sites in the target protein can be identified without time-consuming
assignment procedures. We demonstrated the feasibility of this approach
by using structurally well-characterized interactions between mitogen-activated
protein (MAP) kinase p38α and its inhibitor 2-amino-3-benzyloxypyridine.
Furthermore, we confirmed the recently proposed fatty acid binding
to p38α and confirmed the fatty acid-binding site in the MAP
kinase insert region
Interaction Analysis of FABP4 Inhibitors by X‑ray Crystallography and Fragment Molecular Orbital Analysis
X-ray crystal structural determination
of FABP4 in complex with
four inhibitors revealed the complex binding modes, and the resulting
observations led to improvement of the inhibitory potency of FABP4
inhibitors. However, the detailed structure–activity relationship
(SAR) could not be explained from these structural observations. For
a more detailed understanding of the interactions between FABP4 and
inhibitors, fragment molecular orbital analyses were performed. These
analyses revealed that the total interfragment interaction energies
of FABP4 and each inhibitor correlated with the ranking of the <i>K</i><sub>i</sub> value for the four inhibitors. Furthermore,
interactions between each inhibitor and amino acid residues in FABP4
were identified. The oxygen atom of Lys58 in FABP4 was found to be
very important for strong interactions with FABP4. These results might
provide useful information for the development of novel potent FABP4
inhibitors