Department of Biomedical Engineering, and Chemistry of Life Processes Institute, Northwestern University, Evanston, IL 60208, USA.
Proc Natl Acad Sci U S A. 2013 Feb 26;110(9):3363-8. doi: 10.1073/pnas.1212909110. Epub 2013 Feb 12.
The molecular structure of the yeast nuclear pore complex (NPC) and the translocation of model particles have been studied with a molecular theory that accounts for the geometry of the pore and the sequence and anchoring position of the unfolded domains of the nucleoporin proteins (the FG-Nups), which control selective transport through the pore. The theory explicitly models the electrostatic, hydrophobic, steric, conformational, and acid-base properties of the FG-Nups. The electrostatic potential within the pore, which arises from the specific charge distribution of the FG-Nups, is predicted to be negative close to pore walls and positive along the pore axis. The positive electrostatic potential facilitates the translocation of negatively charged particles, and the free energy barrier for translocation decreases for increasing particle hydrophobicity. These results agree with the experimental observation that transport receptors that form complexes with hydrophilic/neutral or positively charged proteins to transport them through the NPC are both hydrophobic and strongly negatively charged. The molecular theory shows that the effects of electrostatic and hydrophobic interactions on the translocating potential are cooperative and nonequivalent due to the interaction-dependent reorganization of the FG-Nups in the presence of the translocating particle. The combination of electrostatic and hydrophobic interactions can give rise to complex translocation potentials displaying a combination of wells and barriers, in contrast to the simple barrier potential observed for a hydrophilic/neutral translocating particle. This work demonstrates the importance of explicitly considering the amino acid sequence and hydrophobic, electrostatic, and steric interactions in understanding the translocation through the NPC.
酵母核孔复合物(NPC)的分子结构和模型颗粒的易位已通过一种分子理论进行了研究,该理论考虑了孔的几何形状以及核孔蛋白(FG-Nups)的未折叠结构域的序列和锚固位置,这些结构域控制着通过孔的选择性运输。该理论明确地对 FG-Nups 的静电、疏水性、空间位阻、构象和酸碱特性进行建模。由于 FG-Nups 的特定电荷分布,在孔内产生的静电势在靠近孔壁处为负,而在孔轴上为正。正静电势有利于带负电荷颗粒的易位,并且随着颗粒疏水性的增加,易位的自由能障碍减小。这些结果与实验观察结果一致,即与亲水性/中性或带正电荷的蛋白质形成复合物以将其运输通过 NPC 的运输受体既具有疏水性又具有强负电性。分子理论表明,由于在易位颗粒存在下 FG-Nups 的相互依赖的重组,静电和疏水力相互作用对易位势的影响是协作且不等效的。静电和疏水力相互作用的组合会产生复杂的易位势,显示出阱和势垒的组合,而与亲水/中性易位颗粒观察到的简单势垒不同。这项工作表明,在理解 NPC 中的易位时,明确考虑氨基酸序列以及疏水性、静电和空间位阻相互作用非常重要。