Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA.
Nat Chem Biol. 2013 Mar;9(3):169-76. doi: 10.1038/nchembio.1163. Epub 2013 Jan 20.
The ability to chemically control protein-protein interactions would allow the interrogation of dynamic cellular processes and lead to a better understanding and exploitation of self-assembling protein architectures. Here we introduce a new engineering strategy--reverse metal-templated interface redesign (rMeTIR)--that transforms a natural protein-protein interface into one that only engages in selective response to a metal ion. We have applied rMeTIR to render the self-assembly of the cage-like protein ferritin controllable by divalent copper binding, which has allowed the study of the structure and stability of the isolated ferritin monomer, the demonstration of the primary role of conserved hydrogen-bonding interactions in providing geometric specificity for cage assembly and the uniform chemical modification of the cage interior under physiological conditions. Notably, copper acts as a structural template for ferritin assembly in a manner that is highly reminiscent of RNA sequences that template virus capsid formation.
化学控制蛋白质-蛋白质相互作用的能力将允许对动态细胞过程进行询问，并导致对自组装蛋白质结构更好的理解和利用。在这里，我们引入了一种新的工程策略——反向金属模板界面重新设计（rMeTIR）——它将天然蛋白质-蛋白质界面转化为仅对金属离子选择性反应的界面。我们已经应用 rMeTIR 使笼状蛋白铁蛋白的自组装能够通过二价铜结合来控制，这使得我们能够研究分离的铁蛋白单体的结构和稳定性，证明保守氢键相互作用在为笼组装提供几何特异性方面的主要作用，并在生理条件下对笼内部进行均匀的化学修饰。值得注意的是，铜以一种高度类似于模板病毒衣壳形成的 RNA 序列的方式充当铁蛋白组装的结构模板。
