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Supramolecular Control over Split‐Luciferase Complementation
Author(s) -
Bosmans Ralph P. G.,
Briels Jeroen M.,
Milroy LechGustav,
de Greef Tom F. A.,
Merkx Maarten,
Brunsveld Luc
Publication year - 2016
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201602807
Subject(s) - luciferase , complementation , supramolecular chemistry , protein fragment complementation assay , chemistry , downregulation and upregulation , biophysics , combinatorial chemistry , biochemistry , biology , crystallography , transfection , mutant , gene , crystal structure
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 cucurbit[8]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.