A dsDNA-lighted fluorophore for monitoring protein-ligand interaction through binding-mediated DNA protection

• 来源期刊: 中国科学:化学（英文版）
• 年，卷（期）: 2018,（12）
• 页码范围: 1630-1636
• 页数: 7页
• 语种: 英文

摘要

Because of their important roles in cellular functions,life activities,drug screening,and disease treatment,the development of efficient methods for monitoring protein-ligand interaction is essential.In this study,inspired by our previous studies on DNA conformation-selective fluorescent indicators,we developed a new sensing platform for monitoring protein-ligand interaction and detecting protein activity based on binding-mediated DNA protection and the dsDNA-lighted fluorophore,ethyl-4-[3,6-bis (l-methyl-4-vinylpyridium iodine)-9H-carbazol-9-yl)] butanoate (EBCB).The ligand was purposefully linked to the 3'-terminal of a hairpin DNA probe to selectively bind with the target protein and protect the DNA from cleavage by exonuclease Ⅲ.By virtue of EBCB's outstanding capacity to discriminate DNA conformation,the protein-ligand interaction could be effectively monitored through a fluorescence change in EBCB.A high fluorescence signal was detected when the hairpin DNA was protected in the presence of the target protein,whereas a much lower signal was observed in the presence of nontarget proteins.Our results demonstrated that the proposed strategy had high potential,such as high selectivity and relatively high sensitivity,for monitoring protein-ligand interaction and detecting protein activity.We believe these results will pave the way for applying dsDNA-lighted fluorophore EBCB as an effective signal transducer for DNA conformation transformation-mediated biochemical sensing.