Figure 6
From: Dodecin as carrier protein for immunizations and bioengineering applications
Modification of mtDod-mmACP, mtDod-mmACP-H8 and mmACP by Sfp with fluorescent CoA. L: ladder. NC: negative control reaction without Sfp. Eq.: molar equivalents of mmACP loaded onto the SDS-PAGE gel. (a) Top: Coomassie stained SDS-PAGE gel of the reaction solution and negative controls. Sfp, runs at an apparent molecular weight of slightly above 25 kDa, mtDod-mmACP-H8 slightly below 25 kDa and mmACP slightly below 15 kDa. Unmodified mmACP and mtDod-mmACP-H8 (negative controls) show lower apparent molecular weights indicating a different running behaviour between CoA-488 modified and unmodified proteins. Bottom: In-gel fluorescence taken before Coomassie staining. Only proteins modified with the CoA-488 are visible. Higher bands represent labelled mtDod-mmACP-H8 and lower bands labelled mmACP. (b) Top: Coomassie stained SDS-PAGE gel of the reaction solutions of mtDod-mmACP-H8 and mtDod-mmACP after 1 h and 2 h. Bottom: in-gel fluorescence taken before Coomassie staining. For both constructs, fluorescence intensity increases at longer reaction times. For uncropped images and mtDod-mmACP blots see Supplementary Fig. S9.
