Distinct features of InlP as compared to Lmo2027.

<p>(A) Pairwise sequence alignment of InlP and Lmo2027 with conserved amino acids highlighted in red. The LRRs and calcium binding site on InlP are noted. Arrows signify ß-sheets and coils correspond to α-helices. (B) Loading control of pull down experiments on Lmo2027 by Coomassie blue staining. GST protein alone (-), Lmo2027-GST fusion protein (2027) or InlP-GST fusion protein (InlP) bound to glutathione-Sepharose resin used as bait for pull-down experiments with protein extracts from MDCK cells. The elution fraction from pull-down samples using InlP-GST (InlP) as bait were sequentially diluted 5-fold before SDS analysis; undiluted elution fractions were analyzed from pull-down samples using GST (-) or Lmo2027 (2027). Data shown are Coomassie staining and the most abundant band in each lane represents the bait. (C) GST fusion proteins or GST protein alone (-) bound to glutathione-Sepharose resin were incubated overnight with protein extracts from MDCK cells, and elution fractions were analyzed by Western blot with anti-afadin antibodies. The following GST fusion proteins were used: GST-Lmo2027 (Lmo2027) and GST-InlP (InlP). Undiluted elution fractions were analyzed with the exception of the elution fractions marked by black triangles: elution fractions from the experiments using GST-InlP as bait were sequentially diluted by 5-fold before western blot analysis. Actin: loading control. (D) Crystal structure of InlP with schematic depiction of domain layout below. (E) Crystal structure of Lmo2027 with color-coded domains, like in (D).