FIGURE 1.

Identification of CS-E-binding proteins from mouse lung. A, a lung homogenate from a 7-week-old mouse was applied to the CS-E-immobilized column, which was washed stepwise with a buffer containing 0.15 m NaCl followed by those containing 0.5, 1.0, and 2.0 m NaCl. The 0.5 m NaCl eluate was separated by 12% SDS-PAGE and visualized with Coomassie Brilliant Blue. Three protein bands at 46, 52, and 80 kDa were detected (arrowheads a–c). B, shown is the delayed extraction MALDI-TOF mass spectrum of the tryptic peptides obtained from band c in A. Numbered signals were used for a data base search using the MASCOT program. Asterisks indicate the peptide signals from trypsin itself. C, MS signals matched to the peptides derived from RAGE. The numbered underlines correspond to the respective MS signals in B. D, shown is Western blotting using anti-RAGE antibody of the lung homogenate (30 μg as total protein) and 0.15–1.0 m NaCl-eluted fractions (1 μg as total protein) recovered from the CS-E immobilized column.