Figure 3

Gelatin zymography of pollen diffusates and sensitivity to inhibitors. Approximately 10 μg of Kentucky blue grass (lanes 2–6) rye grass (lanes 7–11) and Bermuda grass (lanes 12–16) diffusates were separated on 12.5% SDS/PAGE. Trypsin (~500 ng) was loaded in lane 1. Diffusates (lanes 2, 7 and 12) contain several high MW bands (~95,000) with proteolytic activity and Bermuda grass diffusate also contains intense proteolytic activity at ~35,000 (lane 12). PMSF partially inhibited some of the activity in rye grass (lane 8) but not Kentucky or Bermuda (lanes 3 and 13); complete protease inhibitors (lanes 4, 9 and 14) totally blocked activity; and AEBSF (lanes 5, 10 and 15) was a moderately effective inhibitor. Activity was unaffected by iodoacetamide (lanes 6, 11 and 16).