National Natural Science Foundation of China (NSFC)
31670749
中国
National Natural Science Foundation of China (NSFC)
31925024
中国
Chinese Academy of Sciences
ZDBS-LY-SM003-02; XDB08020302
中国
引用
ジャーナル: Nat Commun / 年: 2021 タイトル: Phospholipid translocation captured in a bifunctional membrane protein MprF. 著者: Danfeng Song / Haizhan Jiao / Zhenfeng Liu / 要旨: As a large family of membrane proteins crucial for bacterial physiology and virulence, the Multiple Peptide Resistance Factors (MprFs) utilize two separate domains to synthesize and translocate ...As a large family of membrane proteins crucial for bacterial physiology and virulence, the Multiple Peptide Resistance Factors (MprFs) utilize two separate domains to synthesize and translocate aminoacyl phospholipids to the outer leaflets of bacterial membranes. The function of MprFs enables Staphylococcus aureus and other pathogenic bacteria to acquire resistance to daptomycin and cationic antimicrobial peptides. Here we present cryo-electron microscopy structures of MprF homodimer from Rhizobium tropici (RtMprF) at two different states in complex with lysyl-phosphatidylglycerol (LysPG). RtMprF contains a membrane-embedded lipid-flippase domain with two deep cavities opening toward the inner and outer leaflets of the membrane respectively. Intriguingly, a hook-shaped LysPG molecule is trapped inside the inner cavity with its head group bent toward the outer cavity which hosts a second phospholipid-binding site. Moreover, RtMprF exhibits multiple conformational states with the synthase domain adopting distinct positions relative to the flippase domain. Our results provide a detailed framework for understanding the mechanisms of MprF-mediated modification and translocation of phospholipids.
名称: bacterial membrane protein / タイプ: CELL 詳細: A recombinant protein from Rhizobium tropici expressed in Escherichia coli cells Entity ID: #1 / 由来: NATURAL
由来(天然)
生物種: Rhizobium tropici (根粒菌) / 細胞内の位置: Membrane
由来(組換発現)
生物種: Escherichia coli (大腸菌)
緩衝液
pH: 7.5
緩衝液成分
ID
濃度
名称
式
Buffer-ID
1
20mM
buffer
Tris-HClトリスヒドロキシメチルアミノメタン
1
2
300mM
salt
NaCl塩化ナトリウム
1
試料
濃度: 6 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES / 詳細: The protein is reconstituted in lipid nanodiscs