Fluorescence resonance energy transfer analysis of protein translocase -: SecYE from Thermus thermophilus HB8 forms a constitutive oligomer in membranes
Fluorescence resonance energy transfer analysis of protein translocase -: SecYE from Thermus thermophilus HB8 forms a constitutive oligomer in membranes
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DOI：
10.1074/jbc.m300230200
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发表时间：
2003-04-18
影响因子：
4.8
通讯作者：
Ito, K
中科院分区：
文献类型：
--
作者：
Mori, H;Tsukazaki, T;Ito, K
SecY和SecE是两个主要的转位酶亚基，其产生用于前蛋白穿过细菌细胞质膜的转运的通道样途径。在这里，我们报告的克隆，表达和纯化的SecYE复合物（TSecYE）从嗜热菌，嗜热栖热菌HB8。纯化的TSecYE可以重组成在T.嗜热菌SecA（TSecA）依赖的前蛋白易位。在重构期间用供体或受体荧光团标记的TSecYE衍生物混合后，荧光共振能量转移实验证明，脂质双层中的2个或更多个TSecYE单元缔合以形成很大程度上不可交换的低聚结构。
SecY and SecE are the two principal translocase subunits that create a channel-like pathway for the transit of preprotein across the bacterial cytoplasmic membrane. Here we report the cloning, expression, and purification of the SecYE complex (TSecYE) from a thermophilic bacterium, Thermus thermophilus HB8. Purified TSecYE can be reconstituted into proteoliposomes that function in T. thermophilus SecA (TSecA) dependent preprotein translocation. After the mixing of TSecYE derivatives labeled with either a donor or an acceptor fluorophore during reconstitution, fluorescence resonance energy transfer experiments demonstrated that 2 or more units of TSecYE in the lipid bilayer associate to form a largely non-exchangeable oligomeric structure.