Functional analyzes on an ABC transporter involved in the selective release of outer membrane lipoproteins from membranes

参与从膜选择性释放外膜脂蛋白的 ABC 转运蛋白的功能分析

• 批准号: 12660068
• 负责人: MATSUYAMA Shin-ichi
• 金额: $ 2.24万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12660068/
• 关键词: lipoprotein; membrane localization; Lol system; ABC transporter; outer membrane; Escherichia coli; ATP; sorting signal; 細胞表層; ABCトフンスボーター; タンパク質局在化; リポ蛋白質; タンパク質輸送; 局在化; 相互作用

We identified a unique ABC transporter, LolCDE, which is responsible for the localization of lipoproteins from the inner to the outer membrane. The ATPase activity of LolCDE was stimulated in an outer membrane lipoprotein-dependent manner, indicating that LolCDE directly recognizes the sorting signal of lipoproteins. The same conclusion was obtained from a genetic analysis on a mutant in which lipoproteins possessing inner membrane sorting signals are localized to the outer membrane. We proposed that the inner membrane sorting signal acts as an avoidance signal on the basis of that inner membrane lipoproteins are not recognized by LolCDE. We showed that N-acylation of the cysteine residue is essential for the Lol system-dependent localization of lipoproteins to the outer membrane. Biochemical analyzes of a mutant LolA protein revealed that LolA directly interacts with LolCDE to transfer lipoproteins. The LolCDE gene as well as lolA and lolB was found to be essential for cell growth of E.coli, and almost all of Gram-negative bacteria have their homologs, suggesting that the membrane localization of lipoproteins by the Lol system is a common feature in Gram-negative bacteria.

我们发现了一种独特的 ABC 转运蛋白 LolCDE，它负责脂蛋白从内膜到外膜的定位。 LolCDE的ATP酶活性以外膜脂蛋白依赖性方式受到刺激，表明LolCDE直接识别脂蛋白的分选信号。从突变体的遗传分析中得到了相同的结论，其中具有内膜分选信号的脂蛋白定位于外膜。基于内膜脂蛋白不被LolCDE识别，我们提出内膜分选信号充当回避信号。我们发现半胱氨酸残基的 N-酰化对于 Lol 系统依赖性脂蛋白定位到外膜至关重要。对突变型 LolA 蛋白的生化分析表明，LolA 直接与 LolCDE 相互作用以转移脂蛋白。 LolCDE 基因以及 lolA 和 lolB 被发现对于大肠杆菌的细胞生长至关重要，并且几乎所有革兰氏阴性细菌都有其同源物，这表明 Lol 系统对脂蛋白的膜定位是革兰氏阴性细菌的共同特征。

项目成果

Narita, S.: "Disruption of lolODE encoding an ATP-binding cassette transporter is lethal for Escherichia coli and prevents the release of lipoproteins from the inner membrane"Journal of Bacteriology. 184. 1417-1422 (2001)

Narita, S.：“编码 ATP 结合盒转运蛋白的 lolODE 的破坏对大肠杆菌来说是致命的，并阻止脂蛋白从内膜释放”《细菌学杂志》。

Yakushi,T.: "A new ABC transporter mediating the detachment of lipid-modified proteins from membranes."Nature Cell Biology. 2. 212-218 (2000)

Yakushi,T.：“一种新的 ABC 转运蛋白介导脂质修饰蛋白从膜上分离。”《自然细胞生物学》。

Shin-ichi Matsuyama: "ATPase and GTPase activities copurifying with GTP-binding proteins in Escherichia coli"Journal of Molecular Microbiology and Biotechnology. 2・1. 261-263 (2000)

Shin-ichi Matsuyama：“大肠杆菌中与 GTP 结合蛋白共纯化的 ATP 酶和 GTP 酶活性”《分子微生物学和生物技术杂志》2・1（2000 年）。

Tanaka, K.: "Deletion of lolB encoding an outer membrane lipoprotein, is lethal for Escherichia coli and causes accumulation of lipoprotein localization intermediates in the periplasm"J.Bacteriol.. 183. 6538-6542 (2001)

Tanaka, K.：“编码外膜脂蛋白的 lolB 的缺失对大肠杆菌是致命的，并导致周质中脂蛋白定位中间体的积累”J.Bacteriol.. 183. 6538-6542 (2001)

Yakushi, T.: "A new ABC transporter mediating the detachment of lipid-modified proteins from membranes."Nature Cell Biol.. 2. 212-218 (2000)

Yakushi, T.：“一种新的 ABC 转运蛋白介导脂质修饰蛋白与膜的分离。”Nature Cell Biol.. 2. 212-218 (2000)