Active Plasma Membrane P-type H+-ATPase Reconstituted into Nanodiscs Is a Monomer
Active Plasma Membrane P-type H+-ATPase Reconstituted into Nanodiscs Is a Monomer
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DOI：
10.1074/jbc.m112.446948
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发表时间：
2013-09-13
影响因子：
4.8
通讯作者：
Fuglsang, Anja Thoe
中科院分区：
文献类型：
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作者：
Justesen, Bo Hojen;Hansen, Randi Westh;Fuglsang, Anja Thoe
质膜H+- atp酶是p型atp酶的一个亚家族，负责将质子泵出细胞，在植物和真菌中对建立和维持关键的跨膜质子梯度至关重要。在这里，我们报道了拟南芥质膜H+- atp酶异构体2重组成可溶性纳米级脂质双层，也称为纳米圆盘。基于天然凝胶分析和交联研究，泵作为功能单体插入纳米圆盘。将H+- atp酶插入纳米圆盘中，可以使用通常只适用于可溶性蛋白质的技术进行结构和功能表征。
Plasma membrane H+-ATPases form a subfamily of P-type ATPases responsible for pumping protons out of cells and are essential for establishing and maintaining the crucial transmembrane proton gradient in plants and fungi. Here, we report the reconstitution of the Arabidopsis thaliana plasma membrane H+-ATPase isoform 2 into soluble nanoscale lipid bilayers, also termed nanodiscs. Based on native gel analysis and cross-linking studies, the pump inserts into nanodiscs as a functional monomer. Insertion of the H+-ATPase into nanodiscs has the potential to enable structural and functional characterization using techniques normally applicable only for soluble proteins.