Nanoscale stiffness topography reveals structure and mechanics of the transport barrier in intact nuclear pore complexes.
Nanoscale stiffness topography reveals structure and mechanics of the transport barrier in intact nuclear pore complexes.
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DOI：
10.1038/nnano.2014.262
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发表时间：
2015-01
影响因子：
38.3
通讯作者：
中科院分区：
文献类型：
--
作者：
核孔复合体（NPC）是细胞核和细胞质之间转运的大门。小分子通过被动扩散穿过NPC，但大于约5nm的分子必须与核转运受体结合以克服NPC内的选择性屏障。虽然NPC的胞质环的结构和形状相对较好地表征，但选择性屏障位于NPC的中央通道深处，并且严重依赖于非结构化的核孔蛋白，因此尚未得到很好的理解。在这里，我们表明，刚度地形与尖锐的原子力显微镜提示可以产生纳米级的NPC横截面。横截面揭示了两种不同的结构，一个细胞质环和一个中央塞结构，这是一致的三维NPC结构来自电子显微镜。在运输循环重新激活和由此产生的货物释放后，中心堵塞物仍然存在，表明堵塞物是NPC屏障的固有部分。添加的核转运受体在完整的转运屏障上积累，并导致屏障刚度的均匀化。在NPC中观察到的纳米力学性质表明存在内聚屏障来运输，并且与NPC通道中核孔蛋白的中心冷凝物的存在定量一致。
The nuclear pore complex (NPC) is the gate for transport between the cell nucleus and the cytoplasm. Small molecules cross the NPC by passive diffusion, but molecules larger than ~5 nm must bind to nuclear transport receptors to overcome a selective barrier within the NPC. Whilst the structure and shape of the cytoplasmic ring of the NPC are relatively well characterized, the selective barrier is situated deep within the central channel of the NPC and depends critically on unstructured nuclear pore proteins,, and is therefore not well understood. Here, we show that stiffness topography with sharp atomic force microscopy tips can generate nanoscale cross sections of the NPC. The cross sections reveal two distinct structures, a cytoplasmic ring and a central plug structure, which are consistent with the three-dimensional NPC structure derived from electron microscopy. The central plug persists after reactivation of the transport cycle and resultant cargo release, indicating that the plug is an intrinsic part of the NPC barrier. Added nuclear transport receptors accumulate on the intact transport barrier and lead to a homogenization of the barrier stiffness. The observed nanomechanical properties in the NPC indicate the presence of a cohesive barrier to transport, and are quantitatively consistent with the presence of a central condensate of nuclear pore proteins in the NPC channel.
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影响因子：
3.4
作者：
Roduit, Charles;Sekatski, Serguei;Kasas, Sandor
通讯作者：
Kasas, Sandor
影响因子：
7
作者：
Yamada, Justin;Phillips, Joshua L.;Rexach, Michael F.
通讯作者：
Rexach, Michael F.
DOI：
10.1039/c1ib00041a
发表时间：
2012-01
期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
--
作者：
Goryaynov A;Ma J;Yang W
通讯作者：
Yang W
影响因子：
64.5
作者：
Adams RL;Wente SR
通讯作者：
Wente SR
影响因子：
9.8
作者：
Zaitseva L;Myers R;Fassati A
通讯作者：
Fassati A