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. Author manuscript; available in PMC: 2013 Jan 23.
Published in final edited form as: Science. 2012 Nov 8;338(6111):1214–1217. doi: 10.1126/science.1228984

Fig. 3. LPS transport can be reconstituted in vitro.

Fig. 3

(A) LPS accumulates in LptC in membrane vesicles over time in an LptBFG and ATP-dependent manner. RSO membrane vesicles containing overexpressed LptC(T47pBPA) with or without co-overexpression of LptBFG were prepared in the presence or absence of ATP. After incubation at 30°C for time indicated, the vesicles were UV-irradiated. Crosslinking was detected as in Fig. 1B. (B) LPS is released to LptA in an LptBFGC, time, and ATP-dependent manner. RSO membrane vesicles were prepared from wild-type cells or cells overexpressing LptC, LptBFG, or LptBFGC with or without added ATP. Purified LptA(I36pBPA or H37pBPA) was added to these vesicles, incubated at 30°C for time indicated, and then UV-irradiated. (C) Model of the stacked crystal structures of LptC (green) and LptA (purple). Residues I36 and H37 in LptA, which interact with LPS and LptC, respectively, are depicted as stick structures. (D) Periplasmic bridge components properly assemble in vitro. Purified LptA(I36pBPA or H37pBPA) was added to LptC- or LptBFGC-enriched RSO membrane vesicles. Samples were incubated at 30°C, and UV-irradiated. LptA(H37pBPA) crosslinked to LptC in an ATP, time, and LptBFG-independent manner.