Selection of Protein-Protein Interactions of Desired Affinities with a Bandpass Circuit.
Selection of Protein-Protein Interactions of Desired Affinities with a Bandpass Circuit.
复制标题
使用带通电路选择所需亲和力的蛋白质-蛋白质相互作用。
DOI：
10.1016/j.jmb.2018.11.011
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发表时间：
2019
影响因子：
5.6
通讯作者：
Woolley,GAndrew
中科院分区：
文献类型：
--
作者：
Brechun,KatherineE;Arndt,KatjaM;Woolley,GAndrew
我们已经在大肠杆菌中开发了一种遗传电路，可以通过改变培养基中抗生素的浓度来选择不同强度的蛋白质-蛋白质相互作用。遗传电路将蛋白质-蛋白质相互作用强度与β-内酰胺酶活性联系起来，同时对β-内酰胺酶活性施加可调的正选择压力和负选择压力。只有当细胞表达的相互作用蛋白质的亲和力落在设定的高通和低通阈值内时，细胞才能存活;即电路因此充当蛋白质-蛋白质相互作用的带通滤波器。我们表明，该电路可用于从具有一系列亲和力的混合群体中恢复所需亲和力的蛋白质-蛋白质相互作用。该电路还可用于选择具有确定强度的蛋白质-蛋白质相互作用的抑制剂。
We have developed a genetic circuit inEscherichia colithat can be used to select for protein–protein interactions of different strengths by changing antibiotic concentrations in the media. The genetic circuit links protein–protein interaction strength to β-lactamase activity while simultaneously imposing tuneable positive and negative selection pressure for β-lactamase activity. Cells only survive if they express interacting proteins with affinities that fall within set high- and low-pass thresholds; i.e. the circuit therefore acts as a bandpass filter for protein–protein interactions. We show that the circuit can be used to recover protein–protein interactions of desired affinity from a mixed population with a range of affinities. The circuit can also be used to select for inhibitors of protein–protein interactions of defined strength.