The DedA superfamily member PetA is required for the transbilayer distribution of phosphatidylethanolamine in bacterial membranes.
The DedA superfamily member PetA is required for the transbilayer distribution of phosphatidylethanolamine in bacterial membranes.
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DOI：
10.1073/pnas.2301979120
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发表时间：
2023-05-16
影响因子：
11.1
通讯作者：
Rudner, David Z.
中科院分区：
文献类型：
--
作者：
Roney, Ian J.;Rudner, David Z.
磷脂是如何在生物膜的内部和外部小叶之间分布的，人们知之甚少。虽然在鉴定真核细胞中的转运体方面取得了进展，但细菌中翻转酶的鉴定仍然难以捉摸。在这里，我们提供的证据表明，DedA超家族成员PetA是一种脂质转运体，在枯草芽孢杆菌膜的内外小叶之间分布磷脂酰乙醇胺。我们的工作表明，DedA家族成员的主要作用是在跨膜运输不同的脂质。磷脂在膜双分子层的内外小叶之间的分选是所有生物体的一个基本问题。尽管经过多年的研究，大多数催化细菌磷脂重定向的酶仍然是未知的。近半个世纪前对枯草芽孢杆菌和巨芽孢杆菌的研究表明，新合成的磷脂酰乙醇胺（PE）迅速转移到双层的外层小叶[Rothman & Kennedy， Proc. Natl]。学会科学。[美国文献74,1821-1825(1977)]但假定的PE翻转酶的身份尚未被发现。最近，DedA超家族的成员参与了细菌脂质载体磷酸十一戊烯基的翻转和体外真核磷脂的混乱。在这里，使用抗菌肽duramycin靶向外向PE，我们发现枯草芽孢杆菌细胞缺乏DedA平行PetA（以前的YbfM）增加了对duramycin的抗性。通过表达枯草芽孢杆菌PetA或其他细菌的同源物，恢复对杜拉霉素的敏感性。对杜拉霉素介导的PE合成诱导杀伤的分析表明，PetA是PE有效运输所必需的。最后，使用荧光标记的杜拉霉素，我们证明缺乏PetA的细胞与野生型相比，其外小叶的PE减少。我们得出结论，PetA是长期寻找的PE转运体。这些数据结合其他DedA类似物的生物信息学分析表明，DedA超家族成员的主要作用是通过膜双分子层运输不同的脂质。
How phospholipids get distributed between the inner and outer leaflets of biological membranes is poorly understood. While progress has been made in identifying transporters in eukaryotic cells, the identity of flippases in bacteria has remained elusive. Here, we provide evidence that the DedA superfamily member PetA is a lipid transporter that distributes phosphatidylethanolamine between the inner and outer leaflets of Bacillus subtilis membranes. Our work suggests that the primary role of DedA family members is in transporting distinct lipids across the membrane. The sorting of phospholipids between the inner and outer leaflets of the membrane bilayer is a fundamental problem in all organisms. Despite years of investigation, most of the enzymes that catalyze phospholipid reorientation in bacteria remain unknown. Studies from almost half a century ago in Bacillus subtilis and Bacillus megaterium revealed that newly synthesized phosphatidylethanolamine (PE) is rapidly translocated to the outer leaflet of the bilayer [Rothman & Kennedy, Proc. Natl. Acad. Sci. U.S.A. 74, 1821–1825 (1977)] but the identity of the putative PE flippase has eluded discovery. Recently, members of the DedA superfamily have been implicated in flipping the bacterial lipid carrier undecaprenyl phosphate and in scrambling eukaryotic phospholipids in vitro. Here, using the antimicrobial peptide duramycin that targets outward-facing PE, we show that Bacillus subtilis cells lacking the DedA paralog PetA (formerly YbfM) have increased resistance to duramycin. Sensitivity to duramycin is restored by expression of B. subtilis PetA or homologs from other bacteria. Analysis of duramycin-mediated killing upon induction of PE synthesis indicates that PetA is required for efficient PE transport. Finally, using fluorescently labeled duramycin we demonstrate that cells lacking PetA have reduced PE in their outer leaflet compared to wildtype. We conclude that PetA is the long-sought PE transporter. These data combined with bioinformatic analysis of other DedA paralogs argue that the primary role of DedA superfamily members is transporting distinct lipids across the membrane bilayer.
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DOI：
10.1083/jcb.202103105
发表时间：
2021-06-07
期刊：
The Journal of cell biology
影响因子：
--
作者：
Li YE;Wang Y;Du X;Zhang T;Mak HY;Hancock SE;McEwen H;Pandzic E;Whan RM;Aw YC;Lukmantara IE;Yuan Y;Dong X;Don A;Turner N;Qi S;Yang H
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影响因子：
4.8
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DOI：
10.1073/pnas.2015556117
发表时间：
2020-10-27
影响因子：
11.1
作者：
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DOI：
10.1073/pnas.1307010110
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影响因子：
11.1
作者：
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通讯作者：
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影响因子：
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