High-resolution structure of the open NaK channel.

High-resolution structure of the open NaK channel.
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DOI:
10.1038/nsmb.1531
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发表时间:
2009-01
影响因子:
16.8
通讯作者:
--
中科院分区:
生物学1区
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我们报道了来自蜡样芽孢杆菌(B. cereus)的非选择性阳离子通道NaK在1.6 Å分辨率下的晶体结构。该结构显示细胞内门处于开放状态,与先前报道的关闭形式相比,这使得NaK成为唯一一种两种构象的三维结构都已知的通道。通道开放遵循一种保守的机制,即利用一个柔性的甘氨酸残基(门控铰链)使内部螺旋弯曲,这种机制在MthK和大多数其他四聚体阳离子通道中都能看到。此外,我们首次观察到并描述了与通道门控相关的不同的亚基间和亚基内重排以及内部螺旋扭转运动。此外,我们在通道孔腔内部更深处鉴定出一个残基Phe92,它可能在开放的孔内形成一个收缩点,限制离子通过通道的通量。通过86Rb通量测定发现,将该残基突变为Ala会导致离子传导速率随后增加。NaK通道开放和关闭两种构象的结构分别与等效的K⁺通道构象(即MthK和KcsA)的结构相关性非常好。
We report the crystal structure of the non-selective cation channel NaK from b. cereus at a resolution of 1.6 Å. The structure reveals the intracellular gate in an open state compared to the closed form reported previously, making NaK the only channel for which the three-dimensional structures of both conformations are known. Channel opening follows a conserved mechanism of inner helix bending utilizing a flexible glycine residue, the gating hinge, seen in MthK and most other tetrameric cation channels. Additionally, distinct inter and intra-subunit rearrangements involved in channel gating are seen and characterized for the first time along with inner helix twisting motions. Furthermore, we identify a residue deeper within the cavity of the channel pore, Phe92, which likely forms a constriction point within the open pore, restricting ion flux through the channel. Mutating this residue to Ala causes a subsequent increase in ion conduction rates as measured by 86Rb flux assays. The structures of both the open and closed conformations of the NaK channel correlate very well with those of equivalent K+ channel conformations, namely MthK and KcsA, respectively.
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