Fig. 6

Effect of the pepsin, Helicobacter pylori protease, and phospholipase C (PLC) on mucin binding protein (MBP) of rat gastric mucosa. The individual rat stomachs were rinsed with buffered saline, bisected to form identical halves of the organ, and subjected to extraction with saline or 1% Triton X-100, incubation with or without protease inhibitors, and incubation with the medium-derived from H. pylori culture (11). (A) Lane 1, MBP-mucin complex extracted from the gastric mucosa, not extracted with Triton X-100, and incubated in the presence of protease inhibitors. Lanes 2 and 3, molecular weight markers. Lanes 4–7, the preparations of the MBP from the stomach halves treated with 1% Triton X-100 and incubated without protease inhibitors in saline (lanes 4, 6) and with medium from H. pylori culture (lanes 5,7). (B) Effect of PLC on the partition of MBP to Triton X-114. Lanes 1 and 3, Triton X-114 phase from the MBP before and after PLC treatment, respectively. Lane 2, water phase from the partition of PLC-treated MBP. The gel in panel A was stained with Coomassie stain. Panel B represents the Western blot containing mucin (Muc) and main binding protein stained with ImmunBlot kit for glycoprotein detection from Bio-Rad (Richmond, CA).