PP-PLA2-mediated hydrolysis of HDL.
A, NDGGE of intact HDL (lane 1) and PP-PLA2-HDL (lane 2). B, SEC chromatograms of intact HDL and PP-PLA2-HDL. C, SDS-PAGE of intact HDL (lane 1) and PP-PLA2-HDL (lane 2). D, NDGGE of guanidine-treated HDL. Lane 1′, Gd-HDL; lane 2′, Gd-PP-PLA2-HDL. E, SEC chromatograms of Gd-treated HDL. F, SDS-PAGE of Gd-HDL (lane 1′) and Gd-PP-PLA2-HDL (lane 2′). For both NDGGE and SDS-PAGE, 5 μg of protein was loaded per lane. Molecular weight standards are the High Molecular Weight Calibration Kit (GE Healthcare) for NDGGE and Precision Plus Protein Dual Xtra standards (Bio-Rad) for SDS-PAGE, respectively. For SEC analysis, samples were loaded at 2 mg/ml (total protein concentration) on a preparative grade Superdex 200 XK 16/100 column and eluted with 10 mm PBS, pH 7.5, at a flow rate of 1 ml/min.