Supramolecular Control over Split‐Luciferase Complementation |
| |
| Authors: | Dr Ralph P G Bosmans Jeroen M Briels Dr Lech‐Gustav Milroy Dr Tom F A de?Greef Prof Maarten Merkx Prof Luc Brunsveld |
| |
| Affiliation: | Laboratory of Chemical Biology and Institute of Complex Molecular Systems, Department of Biomedical Engineering, Eindhoven University of Technology, AZ, Eindhoven, The Netherlands |
| |
| Abstract: | Supramolecular split‐enzyme complementation restores enzymatic activity and allows for on–off switching. Split‐luciferase fragment pairs were provided with an N‐terminal FGG sequence and screened for complementation through host‐guest binding to cucurbit8]uril (Q8). Split‐luciferase heterocomplex formation was induced in a Q8 concentration dependent manner, resulting in a 20‐fold upregulation of luciferase activity. Supramolecular split‐luciferase complementation was fully reversible, as revealed by using two types of Q8 inhibitors. Competition studies with the weak‐binding FGG peptide revealed a 300‐fold enhanced stability for the formation of the ternary heterocomplex compared to binding of two of the same fragments to Q8. Stochiometric binding by the potent inhibitor memantine could be used for repeated cycling of luciferase activation and deactivation in conjunction with Q8, providing a versatile module for in vitro supramolecular signaling networks. |
| |
| Keywords: | cooperativity cucurbit[8]uril split-luciferase supramolecular chemical biology switching |
|
|
