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. Author manuscript; available in PMC: 2012 Jul 13.
Published in final edited form as: Biol Chem. 2010 Jun;391(6):705–716. doi: 10.1515/BC.2010.066

Figure 7. Effect of purified aeruginolysin on transglutaminase-mediated crosslinking of elafin to fibronectin.

Figure 7

Recombinant elafin (100 ng, 3 μM) was incubated for 2h at 37°C with increasing amounts of aeruginolysin (lanes 1 and 2: no enzyme, 3 and 4: 1 nM, 5 and 6: 10 nM, 7 and 8: 100 nM, 9 and 10: 1000 nM). Samples were treated with GM6001 to inactivate aeruginolysin activity and incubated for 1h at 37°C with fibronectin in absence (lanes 1, 3, 5, 7, 9) or in presence (lanes 2, 4, 6, 8, 10) of 0.38 mU guinea pig liver transglutaminase (TGase). Samples were analyzed by SDS-PAGE followed by Western blot using (A) a biotinylated anti-elafin antibody and (B) an anti-fibronectin antibody. In (A), intact and cleaved elafin (respectively denoted elafin and elafin*) are indicated by arrow heads.