Inhibitors of Fatty Acid Synthesis Induce PPARα-Regulated Fatty Acid β-Oxidative Genes: Synergistic Roles of L-FABP and Glucose
Representative binding curves of stearic acid binding to L-FABP (panel a) and SCP2 (panel b). NBD-stearic acid binding curves were obtained as described in Methods. Briefly, a 2 mL sample of L-FABP (25 nM) or SCP-2 (25 nM) in 10 mM phosphate buffer (pH 7.4) was titrated with small increments of NBD stearate at 24°C (Methods). NBD-stearate fluorescence emission spectra (515–600 nm) were recorded with 490 nm excitation. : fluorescence intensity of NBD stearate in the presence of proteins (at 530 nm for binding to SCP-2 and at 548 nm for binding to L-FABP) and being NBD-stearate fluorescence intensity in buffer (at the same wavelength as for ). Insets are double reciprocal plots of the fluorescence binding data in the same panel.