Covalent attachment of active enzymes to upconversion phosphors allows ratiometric detection of substrates

Burgess, Letitia; Wilson, Hannah; Jones, Alex R.; Harvey, Peter; Natrajan, Louise; Hay, Sam

Abstract

Upconverting phosphors (UCPs) convert multiple low energy photons into higher energy emission via the process of photon upconversion and offer an attractive alternative to organic fluorophores for use as luminescent probes. Here, UCPs were capped with functionalized silica in order to provide a surface to covalently conjugate proteins with surface?accessible cysteines. Variants of green fluorescent protein (GFP) and the flavoenzyme pentaerythritol tetranitrate reductase (PETNR) were then attached via maleimide?thiol coupling in order to allow energy transfer from the UCP to the GFP or flavin cofactor of PETNR, respectively. PETNR retains its activity when coupled to the UCPs, which allows reversible detection of enzyme substrates via ratiometric sensing of the enzyme redox state.

• Journal Article Type: Article
• Acceptance Date: May 27, 2020
• Online Publication Date: Oct 16, 2020
• Publication Date: Nov 20, 2020
• Deposit Date: Jul 7, 2020
• Publicly Available Date: Oct 16, 2020
• Journal: Chemistry – A European Journal
• Print ISSN: 0947-6539
• Electronic ISSN: 1521-3765
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 26
• Issue: 65
• Pages: 14817-14822
• DOI: https://doi.org/10.1002/chem.202001974
• Keywords: General Chemistry
• Public URL: https://nottingham-repository.worktribe.com/output/4755165
• Publisher URL: https://chemistry-europe.onlinelibrary.wiley.com/doi/abs/10.1002/chem.202001974