Non-bilayer Lipids Stimulate the Activity of the Reconstituted Bacterial Protein Translocase*

Non-bilayer Lipids Stimulate the Activity of the Reconstituted Bacterial Protein Translocase*
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非双层脂质刺激重组细菌蛋白转位酶的活性*

DOI:
10.1074/jbc.275.4.2472
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发表时间:
2000
期刊:
The Journal of Biological Chemistry
影响因子:
--
通讯作者:
A. Driessen
A. Driessen
中科院分区:
--
文献类型:
--
作者:
C. van der Does;J. Swaving;W. van Klompenburg;A. Driessen

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为了确定前蛋白转位酶在体外的磷脂需求,大肠杆菌SecYEG复合物纯化的脱脂形式使用洗涤剂十二烷基麦芽糖苷。将SecYEG重构成由确定的合成磷脂组成的脂质体,并分析脂蛋白体的前蛋白易位和SecA易位ATP酶活性。活性严格要求存在的阴离子磷脂,而非双层脂质磷脂酰乙醇胺被发现刺激。后者的效果也可以诱导二油酰甘油,采用非双层构象的脂质。磷脂酰乙醇胺衍生物,更喜欢双层状态不能刺激易位。在不存在SecG的情况下,活性降低,但磷脂需求不变。值得注意的是,发现非双层脂质对于枯草芽孢杆菌SecYEG复合物的活性至关重要。最佳活性需要阴离子和非双层脂质的混合物,其浓度对应于在天然膜中发现的浓度。
To determine the phospholipid requirement of the preprotein translocase in vitro, the Escherichia coli SecYEG complex was purified in a delipidated form using the detergent dodecyl maltoside. SecYEG was reconstituted into liposomes composed of defined synthetic phospholipids, and proteoliposomes were analyzed for their preprotein translocation and SecA translocation ATPase activity. The activity strictly required the presence of anionic phospholipids, whereas the non-bilayer lipid phosphatidylethanolamine was found stimulatory. The latter effect could also be induced by dioleoylglycerol, a lipid that adopts a non-bilayer conformation. Phosphatidylethanolamine derivatives that prefer the bilayer state were unable to stimulate translocation. In the absence of SecG, activity was reduced, but the phospholipid requirement was unaltered. Remarkably, non-bilayer lipids were found essential for the activity of the Bacillus subtilis SecYEG complex. Optimal activity required a mixture of anionic and non-bilayer lipids at concentrations that correspond to concentrations found in the natural membrane.
磷脂酰乙醇胺及其N-甲基衍生物的结构和热致性质。
DOI: 10.1021/bi00306a030
发表时间: 1984
期刊: Biochemistry
影响因子: 2.9
作者:
Mulukutla,S;Shipley,GG
通讯作者: Shipley,GG
纯化的大肠杆菌 secB 蛋白可在体外延迟折叠并促进麦芽糖结合蛋白的膜易位。
DOI: 10.1073/pnas.85.23.8978
发表时间: 1988
影响因子: 11.1
作者:
Weiss,JB;Ray,PH;BassfordJr,PJ
通讯作者: BassfordJr,PJ
阴离子磷脂和部分解折叠可促进部分大肠杆菌 SecA 蛋白深入渗透到模型膜中。
DOI: --
发表时间: 1992
期刊: The Journal of biological chemistry
影响因子: --
作者:
Ulbrandt,ND;London,E;Oliver,DB
通讯作者: Oliver,DB