Figure 6 : In vitro binding between Drosophila Sarcoglycans and EGFR.

(A) Diagrams of the obtained constructs for the production of recombinant proteins. The numbers written below the rectangular bars represent the amino acid numbers counted from the N-termini of the respective proteins. (B) Western blots of both Input and Elution samples with anti-His or anti-Flag antibodies are shown. Binding between the extracellular soluble form of Drosophila EGFR (Egfr1-559) and variants of Sarcoglycans was examined. 5 mg of total protein in the E.coli extract was applied for a 100 μl volume of the Ni-NTA agarose gel and eluted with 100 μl buffer. 20 μl of each sample was loaded for the SDS-PAGE. While no binding of Egfr1-559 with dScgα was seen (I-lane 12), dScgβ and dScgδ demonstrated binding to Egfr1-559 (II and IV-lane 12). Furthermore, neither dScgβ nor dScgδ, lacking any EGF-like consensus sequence, showed binding to Egfr1-559 (III and V-lane 12), suggesting that the binding between dScgβ and Egfr1-559, dScgδ and Egfr1-559 is mediated by the EGF-like consensus sequence in the C-terminus regions of dscgβ and dScgδ.

Yamaguchi et al.HOAJ BIOLOGY  2012 1:7DOI : 10.7243/2050-0874-1-7