Location of Pim-1 mutations and spectral properties of wild type (black), Y53H (red), E124Q (green), E135K (pink) and E142D (blue).

<p>(<b>A</b>) Structure of Pim-1 (PDB code: 1XWS) shown as a ribbon diagram. Mutated residues are highlighted and the main secondary structure elements are shown. The ATP mimetic inhibitor cocrystallized in this structure is shown in ball and stick representation.(<b>B</b>) Detailed view of the local structural environment around the mutated residue Y53. (<b>C</b>) Detailed view of the local structural environment around the mutated residue E135. (<b>D</b>) Near-UV CD spectra were recorded in a 1.0-cm quartz cuvette at 1.4 mg/ml protein concentration in 20 mM Tris/HCl, pH 7.5 containing 0.2 M NaCl and 2 mM DTT. (<b>E</b>) Intrinsic fluorescence emission spectra were recorded at 0.04 mg/ml protein concentration (295 nm excitation wavelength) at 10°C in 20 mM Tris/HCl, pH 7.5 containing 0.2 M NaCl and 200 µM DTT. (<b>F</b>) Far-UV CD spectra were recorded in a 0.1-cm quartz cuvette at 0.2 mg/ml in 20 mM Tris/HCl, pH 7.5 containing 0.2 M NaCl and 0.4 mM DTT.