Univ. Bordeaux, IECB, Laboratoire ARNA, 2 rue Robert Escarpit, 33607 Pessac (France); Inserm U869, Laboratoire ARNA, 146 rue Léo Saignat, 33076 Bordeaux (France).
Angew Chem Int Ed Engl. 2014 Jul 1;53(27):6942-5. doi: 10.1002/anie.201400402. Epub 2014 Jun 10.
Biosensors derived from aptamers were designed for which folding into a hairpin shape is triggered by binding of the cognate ligand. These aptamers (termed aptaswitches) thus switch between folded and unfolded states in the presence and absence of the ligand, respectively. The apical loop of the folded aptaswitch is recognized by a second hairpin called the aptakiss through loop-loop or kissing interactions, whereas the aptakiss does not bind the unfolded aptaswitch. Therefore, the formation of a kissing complex signals the presence of the ligand. Aptaswitches were designed that enable the detection of GTP and adenosine in a specific and quantitative manner by surface plasmon resonance when using a grafted aptakiss or in solution by anisotropy measurement with a fluorescently labeled aptakiss. This approach is generic and can potentially be extended to the detection of any molecule for which hairpin aptamers have been identified, as long as the apical loop is not involved in ligand binding.
基于适体的生物传感器被设计用于当配体结合时，折叠成发夹形状。这些适体（称为适体开关）分别在配体存在和不存在的情况下在折叠和未折叠状态之间切换。发夹的顶端环通过环-环或亲吻相互作用被称为适体吻的第二个发夹识别，而适体吻不结合未折叠的适体开关。因此，亲吻复合物的形成表明配体的存在。已经设计了适体开关，当使用接枝的适体吻时，通过表面等离子体共振以特定和定量的方式检测 GTP 和腺苷，或者通过用荧光标记的适体吻进行各向异性测量时，在溶液中检测 GTP 和腺苷。这种方法是通用的，并且可以潜在地扩展到任何已经鉴定出发夹适体的分子的检测，只要顶端环不参与配体结合。
