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Structure paper

タイトル	Structural basis of MsbA-mediated lipopolysaccharide transport.
ジャーナル・号・ページ	Nature, Vol. 549, Issue 7671, Page 233-237, Year 2017
掲載日	2017年9月14日
著者	Wei Mi / Yanyan Li / Sung Hwan Yoon / Robert K Ernst / Thomas Walz / Maofu Liao /
PubMed 要旨	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 ...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.
リンク	Nature / PubMed:28869968 / PubMed Central
手法	EM (単粒子)
解像度	4.2 - 6.9 Å
構造データ	EMDB-8465: 3D cryo-EM reconstruction of MsbA-nanodisc with ADP 手法: EM (単粒子) / 解像度: 6.9 Å 8467 5ttp EMDB-8467, PDB-5ttp: Cryo-EM structure of MsbA-nanodisc with ADP-vanadate 手法: EM (単粒子) / 解像度: 4.8 Å 8469 5tv4 EMDB-8469, PDB-5tv4: 3D cryo-EM reconstruction of nucleotide-free MsbA in lipid nanodisc 手法: EM (単粒子) / 解像度: 4.2 Å EMDB-8669: CryoEM map of MsbA in POPG nanodisc 手法: EM (単粒子) / 解像度: 5.0 Å EMDB-8670: CryoEM map of MsbA with lipid A in POPG nanodisc 手法: EM (単粒子) / 解像度: 4.5 Å EMDB-8671: CryoEM map of MsbA in POPC nanodisc 手法: EM (単粒子) / 解像度: 5.5 Å
化合物	ChemComp-PO4: PHOSPHATE ION / ホスファ-ト ChemComp-FTT: 3-HYDROXY-TETRADECANOIC ACID / (R)-3-ヒドロキシテトラデカン酸 ChemComp-MYR: MYRISTIC ACID / ミリスチン酸 ChemComp-DAO: LAURIC ACID / ラウリン酸
由来	Escherichia coli (大腸菌) escherichia coli o157:h7 (大腸菌)
キーワード	HYDROLASE / ABC transporter / LPS / flippase / nanodisc