Lipids are critical components of membranes that could affect the properties of membrane proteins, yet the precise compositions of lipids surrounding membrane-embedded protein complexes is often difficult to discern. Here we report that, for the heterodimeric ABC transporter TmrAB, the extent of delipidation can be controlled by timed exposure to detergent. We subsequently characterize the cohort of endogenous lipids that are extracted in contact with the membrane protein complex, and show that with prolonged delipidation the number of neutral lipids is reduced in favour of their negatively charged counterparts. We show that lipid A is retained by the transporter and that the extent of its binding decreases during the catalytic cycle, implying that lipid A release is linked to adenosine tri-phosphate hydrolysis. Together, these results enable us to propose that a subset of annular lipids is invariant in composition, with negatively charged lipids binding tightly to TmrAB, and imply a role for this exporter in glycolipid translocation.
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The authors thank C. Schmidt for help with the LC-MS experiments and all the members of C.V.R.'s group for stimulating discussions. The authors also acknowledge funding from European Research Council Integral Membrane Proteins Resolution of Stoichiometry and Structure (ERC IMPRESS), the Royal Society and the Germany Research Foundation (SFB 807 and TA157/7 to R.T.) as well as the European Drug Initiative on Channels and Transporters (EDICT to R.T.) funded by the European Commission Seventh Framework. M.T.D. is supported by the Swiss National Science Foundation.
The authors declare no competing financial interests.
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Bechara, C., Nöll, A., Morgner, N. et al. A subset of annular lipids is linked to the flippase activity of an ABC transporter. Nature Chem 7, 255–262 (2015). https://doi.org/10.1038/nchem.2172
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