Sequential DNA Binding and Dimerization Processes of the Photosensory Protein EL222,Biochemistry

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Sequential DNA Binding and Dimerization Processes of the Photosensory Protein EL222
Biochemistry ( IF 2.9 ) Pub Date : 2018-02-12 00:00:00 , DOI: 10.1021/acs.biochem.7b01206
Akira Takakado ₁ , Yusuke Nakasone ₁ , Masahide Terazima ₁

Affiliation

EL222 is a blue light sensor protein, which consists of a light–oxygen–voltage domain as a light sensor and a LuxR-type helix–turn–helix DNA-binding domain. The reaction dynamics of the protein–DNA binding were observed for the first time using the time-resolved transient grating method. The reaction scheme was determined, showing that photoexcited EL222 first binds DNA and the ground state EL222 monomer is subsequently associated with the complex. Rate constants on the millisecond scale were determined for these processes. In addition, binding rates for EL222 with three DNA sequences, with different binding affinities, were measured. Although EL222 binds nonspecific DNA sequences with affinities at least 5-fold lower than the target sequence affinity, the binding rates were almost the same as that for the target DNA. This observation indicates that the specific and nonspecific binding affinities are mainly controlled by differences in the dissociation of DNA binding.

中文翻译：

感光DNA EL222的顺序DNA结合和二聚化过程

EL222是一种蓝光传感器蛋白，它由作为光传感器的光-氧-电压域和LuxR型螺旋-转角-螺旋DNA结合域组成。使用时间分辨瞬变光栅方法首次观察到了蛋白质与DNA结合的反应动力学。确定反应方案，表明光激发的EL222首先结合DNA，并且基态EL222单体随后与该复合物缔合。对于这些过程，确定了毫秒级的速率常数。另外，测量了EL222与具有不同结合亲和力的三个DNA序列的结合率。尽管EL222以比靶序列亲和力低至少5倍的亲和力结合非特异性DNA序列，但结合率几乎与靶DNA的结合率相同。

更新日期：2018-02-12

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