FIGURE 1.

Purification of human LRG by Cyt c affinity chromatography and CD spectrum of the purified human LRG. A, Cyt c affinity column chromatography of human serum. Human serum was applied to a Cyt c-coupled Hi-Trap NHS-activated HP column, which had been equilibrated with 20 mm phosphate buffer (pH 7.4) containing 0.15 m NaCl. The column was washed with the same buffer, and the adsorbed proteins were eluted with 100 mm acetate buffer (pH 4.0) containing 0.5 m NaCl. SDS-PAGE of the whole serum (lane 1) and the fractions (lane 2, F-1; lane 3, F-II) obtained after Cyt c affinity column chromatography is shown. M, molecular weight markers. LRG was detected by Western blot analysis using a rabbit antiserum raised against human LRG (lane 4). B, CD spectra of human LRG and G. brevicaudus PLIβ. The spectra were measured at 20 °C, pH 7.5, and ionic strength 0.1.