Supramolecular Control over Split‐Luciferase Complementation

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 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.

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Highlights from the 52nd EUCHEM conference on stereochemistry, Bürgenstock, Switzerland, May 2017

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“…5). [47][48][49][50][51] The lecture finished with a collection of selected examples of supramolecular protein assembly triggered by DNA-origami, supramolecular wires of proteins and different discotic self-assembled tubular constructs with functional properties. 52 The first Thursday morning lecture was from Prof. Karl-Heinz Altmann (ETH Zurich) entitled "Total Synthesis and Functional Exploration of Macrocyclic Natural Products".…”