Figure 2.
(A) Averaged fluorescence intensities of a CohE-CBM-ybbR-CoA647-labeled surface functionalized with ybbR-Titin-Ig-LPETGG and ybbR-sfGFP-LPETGG. Each protein was immobilized at two separate spots that were then incubated with either GGG-dockerin and sortase or GGG-dockerin but not with sortase. To test for successful ligation of dockerins, CohE-CBM-ybbR-CoA647 was allowed to bind for 10 min at 300 nM, then rinsed and imaged immediately afterward. Fluorescent intensities of each construct were normalized to the intensity of the sortase-positive spot. (B) SDS-PAGE demonstrating the ligation of GGG-dockerin to ybbR-Titin-LPETGG with wild-type sortase A (wt-Srt), pentamutant sortase A (eSrt), or no sortase as negative control. The red arrows are indicating the ligation products.