Photoregulation of a peptide-RNA interaction on a gold surface
Photoregulation of a peptide-RNA interaction on a gold surface
复制标题
DOI：
10.1021/ja071298x
复制
发表时间：
2007-07-18
影响因子：
15
通讯作者：
Nakatani, Kazuhiko
中科院分区：
文献类型：
--
作者：
Hayashi, Gosuke;Hagihara, Masaki;Nakatani, Kazuhiko
核糖开关的发现加速了对RNA与小分子有机化合物相互作用的研究。对于生物学家来说，通过RNA和小分子的相互作用来人为地、可逆地控制体内基因的表达将是非常重要的。在本文中，我们报道了通过体外筛选获得的RNA适配子，其中含有偶氮苯部分的光敏短肽两侧带有精氨酸残基作为配基，提供了与固定在金表面的配位肽的可逆结合。我们设计并合成了一种能与RNA相互作用、可在光照射下可逆地转化其构象、可大规模生产用于体外筛选的光敏短肽。RNA库包含N70个随机序列，在第八个周期后，我们发现了显示K-d约为几个微摩尔的RNA适配子。表面等离子体共振(SPR)实验表明，在360 nm的光照下，RNA适配体可以与金表面固定的多肽的反式异构体结合，但不能与光异构化的顺式多肽结合。SPR信号在430 nm光照射后恢复，导致多肽从顺式异构化为反式异构化。
The discovery of riboswitching has accelerated research on the interaction between RNA and small organic compounds. It will be important for biologists to artificially and reversibly control gene expression in vivo through the interaction of RNA and small molecules. In this paper, we report that RNA aptamers obtained from in vitro selection in which a photoresponsive short peptide containing the azobenzene moiety with flanking arginine residues on both sides as a ligand provided reversible binding to the ligand peptide immobilized onto the gold surface. We designed and synthesized a photoresponsive short peptide that can interact with RNA, can convert its conformation reversibly by photoirradiation, and can be produced on a large scale for in vitro selection. The RNA pool contained N70 random sequences, and after the eighth cycle, we identified RNA aptamers showing the K-d of about a few micromolar. A surface plasmon resonance (SPR) experiment revealed that RNA aptamers could bind to the trans-isomer of the peptide immobilized on the gold surface but not to the cis-peptide isomerized by photoirradiation with 360 nm light to the gold surface. The SPR signals were recovered after photoirradiation with 430 nm light, leading to isomerization of the peptide from cis to trans.