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. Author manuscript; available in PMC: 2014 Aug 15.
Published in final edited form as: Cell Host Microbe. 2013 Aug 14;14(2):195–206. doi: 10.1016/j.chom.2013.07.012

Figure 2. E-cadherin is FadA receptor.

Figure 2

A. Schematic representation of the E-cadherin (CDH1) structure. E-cadherin has five extracellular cadherin (ECs) repeats, numbered EC1–5 starting from N-terminal. TM, transmembrane domain; C, cytoplasmic domain. B. E-cadherin is expressed in epithelial cell HEK293 and most CRC cells. E-cadherin in HEK293 and human CRC cell lines HCT116, RKO, HT29, SW480 and DLD1 was examined by Western blot. Human umbilical vein endothelial cells (HUVEC) were included as a negative control. The endogenous GAPDH was used as a loading control. C. E-cadherin co-immunoprecipitates with FadAc. HEK293 cell lysate expressing E-cadherin was mixed with E. coli lysates expressing FadAc, or mFadA, or BSA, followed by incubation with mouse anti-CDH1 monoclonal antibodies (mAb) and captured with agarose A/G beads. E-cadherin and FadA in the bead elutes were detected by Western blot. D. FadA binds to EC5. Purified GST or GST-fusion proteins carrying EC1–3, EC4, or EC5 were incubated with E. coli lysates expressing FadAc, followed by capture with GST resin. The eluted components were subjected to SDS-PAGE, followed by Coommassie blue staining (top panel) and Western blot (WB) using anti-FadA mAb 5G11-3G8 (bottom panel). See also Figure S2.