Figure 5: Fluorescence binding curves for DANS-As-labeled E2·E3BP derived lipoyl domains on titration by PDK isoforms.
From: Global view of cognate kinase activation by the human pyruvate dehydrogenase complex
The DANS-As-labeled L1 (1.0 μM), or L2S (1.5 μM) or L1L2S MS-1 (2.5 μM) or L1L2S MS-2 (3.0 μM) in 30 mM KH2PO4 (pH 7.5) was titrated by PDK1 (0.78–37.93 μM) or PDK2 (0.18–4.38 μM) or PDK3 (0.24–3.86 μM) or PDK4 (0.24–4.2 μM) at 25 °C. The excitation wavelength was 338 nm and the emission spectra were recorded in the 425–600 nm range. In all cases the fluorescence intensity of the DANS-labeled lipoyl domains was enhanced on PDKs binding. For experimental details see SI Experimental procedures.
