Crosslinking of a Peritrophic Matrix Protein Protects Gut Epithelia from Bacterial Exotoxins
Fig 2
TG-dependent polymerization of drosocrystallin in vitro and in vivo.
(A) Wild-type drosocrystallin (WT) or the KR mutant (KR) was incubated with TG, and subjected to SDS-PAGE in 10% TGX FastCast gels (Bio-Rad Laboratories). These recombinants were detected by Western blotting with a horseradish peroxidase-conjugated anti-6 × His tag antibody. Monodansylcadaverine (MDC) was used as an inhibitor of protein-protein crosslinking. Data are representative of at least three independent experiments. (B) Gut extracts from systemic TG-RNAi flies (Da>TG IR) and their counterparts (Da>+) were subjected to SDS-PAGE in 12% slab gels. Native drosocrystallin in the extracts was detected by Western blotting with an anti-drosocrystallin antibody (Anti-Dcy). Asterisks indicate unknown cross-reacted proteins. Data are representative of three independent experiments.