Structural and Computational Biology Unit, EMBL, Meyerhofstrasse 1, 69117 Heidelberg (Germany).
Angew Chem Int Ed Engl. 2014 Jul 7;53(28):7364-7. doi: 10.1002/anie.201403694. Epub 2014 Jun 4.
Intrinsically disordered proteins (IDPs) can bind to multiple interaction partners. Numerous binding regions in the IDP that act in concert through complex cooperative effects facilitate such interactions, but complicate studying IDP complexes. To address this challenge we developed a combined fluorescence correlation and time-resolved polarization spectroscopy approach to study the binding properties of the IDP nucleoporin153 (Nup153) to nuclear transport receptors (NTRs). The detection of segmental backbone mobility of Nup153 within the unperturbed complex provided a readout of local, region-specific binding properties that are usually masked in measurements of the whole IDP. The binding affinities of functionally and structurally diverse NTRs to distinct regions of Nup153 can differ by orders of magnitudes-a result with implications for the diversity of transport routes in nucleocytoplasmic transport.
无规卷曲蛋白（IDP）可以与多个相互作用伙伴结合。IDP 中的许多结合区域通过复杂的协同作用协同作用，从而促进这种相互作用，但这也使 IDP 复合物的研究变得复杂。为了解决这个挑战，我们开发了一种组合荧光相关和时间分辨偏振光谱方法来研究无规卷曲蛋白核孔蛋白 153（Nup153）与核转运受体（NTR）的结合特性。在未受干扰的复合物中检测 Nup153 的分段骨架流动性提供了局部、区域特异性结合特性的读数，这些特性通常在整个 IDP 的测量中被掩盖。功能和结构上不同的 NTR 与 Nup153 不同区域的结合亲和力可能相差几个数量级——这一结果对核质转运中运输途径的多样性具有重要意义。
