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Structure paper

TitleStructural basis of MsbA-mediated lipopolysaccharide transport.
Journal, issue, pagesNature, Vol. 549, Issue 7671, Page 233-237, Year 2017
Publish dateSep 14, 2017
AuthorsWei Mi / Yanyan Li / Sung Hwan Yoon / Robert K Ernst / Thomas Walz / Maofu Liao /
PubMed AbstractLipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria is critical for the assembly of their cell envelopes. LPS synthesized in the cytoplasmic leaflet of the inner membrane is ...Lipopolysaccharide (LPS) in the outer membrane of Gram-negative bacteria is critical for the assembly of their cell envelopes. LPS synthesized in the cytoplasmic leaflet of the inner membrane is flipped to the periplasmic leaflet by MsbA, an ATP-binding cassette transporter. Despite substantial efforts, the structural mechanisms underlying MsbA-driven LPS flipping remain elusive. Here we use single-particle cryo-electron microscopy to elucidate the structures of lipid-nanodisc-embedded MsbA in three functional states. The 4.2 Å-resolution structure of the transmembrane domains of nucleotide-free MsbA reveals that LPS binds deep inside MsbA at the height of the periplasmic leaflet, establishing extensive hydrophilic and hydrophobic interactions with MsbA. Two sub-nanometre-resolution structures of MsbA with ADP-vanadate and ADP reveal an unprecedented closed and an inward-facing conformation, respectively. Our study uncovers the structural basis for LPS recognition, delineates the conformational transitions of MsbA to flip LPS, and paves the way for structural characterization of other lipid flippases.
External linksNature / PubMed:28869968 / PubMed Central
MethodsEM (single particle)
Resolution4.2 - 6.9 Å
Structure data

EMDB-8465:
3D cryo-EM reconstruction of MsbA-nanodisc with ADP
Method: EM (single particle) / Resolution: 6.9 Å

8467

5ttp

EMDB-8467, PDB-5ttp:
Cryo-EM structure of MsbA-nanodisc with ADP-vanadate
Method: EM (single particle) / Resolution: 4.8 Å

8469

5tv4

EMDB-8469, PDB-5tv4:
3D cryo-EM reconstruction of nucleotide-free MsbA in lipid nanodisc
Method: EM (single particle) / Resolution: 4.2 Å

EMDB-8669:
CryoEM map of MsbA in POPG nanodisc
Method: EM (single particle) / Resolution: 5.0 Å

EMDB-8670:
CryoEM map of MsbA with lipid A in POPG nanodisc
Method: EM (single particle) / Resolution: 4.5 Å

EMDB-8671:
CryoEM map of MsbA in POPC nanodisc
Method: EM (single particle) / Resolution: 5.5 Å

Chemicals

ChemComp-PO4:
PHOSPHATE ION / Phosphate

ChemComp-FTT:
3-HYDROXY-TETRADECANOIC ACID

ChemComp-MYR:
MYRISTIC ACID / Myristic acid

ChemComp-DAO:
LAURIC ACID / Lauric acid

Source
  • Escherichia coli (E. coli)
  • escherichia coli o157:h7 (bacteria)
KeywordsHYDROLASE / ABC transporter / LPS / flippase / nanodisc

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