A Mycobacterium ESX-1–Secreted Virulence Factor with Unique Requirements for Export (original) (raw)
Figure 5
GST Pulldown Analysis of EspB and Rv3879 Interactions
(A) Agarose beads with immobilized GST, GST-SHP1, GST-syntaxin2, GST-Mh3879c, and GST-EspBM were incubated with lysates of E. coli that express V5-Mh3879c and V5-EspBM.
(B) Agarose beads with immobilized GST, GST-SHP1, GST-syntaxin2, GST-Rv3879c, or GST-EspBT were incubated with lysates of E. coli that express V5-Rv3871, V5-Rv3879c, and V5-EspBT. Proteins from cell lysates retained on the beads after washing were separated by SDS-PAGE and detected by western blotting with an antibody against V5. To the right of each set of pulldowns, 0.1% of the input E. coli lysate was analyzed. EspBM physically interacts with Mh3879c, EspBT physically interacts with Rv3879c, and Rv3879c also interacts with Rv3871.