The development of maleimide derived fluorophores for peptide-based applications

Husband, Jonathan Tristan ORCID: 0000-0003-4056-7477 (2020). The development of maleimide derived fluorophores for peptide-based applications. University of Birmingham. Ph.D.

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Abstract

Substituted maleimide fluorophores have been extensively used to study polymeric systems, as their optical properties are responsive to polymer environment and assembly. They are particularity effective as a result of their small size, and ease of functionalization. Furthermore, this makes them well-suited for peptide-based applications too, where fluorescence labelling is essential for tracking peptide therapeutics and disease diagnostics. Initially in this work, the conjugation of maleimide precursors to enzymes is investigated as a turn-on fluorescence conjugation technique. However, despite the successful effective conjugation, the fluorescence of the conjugates was poor from solvent quenching interactions. Further work focused on improving aqueous fluorescence. Firstly, the dye imide substitution was investigated in an effort to prevent protic solvent quenching events, which led to insights into the fluorescence mechanism including improved solid-state fluorescence through preventing pi-pi interactions. Secondly, the synthesis of fluorescent amino acids was successfully undertook for incorporation deeper into enzymes, where solvent quenching should be minimal. In addition, the synthesis of alkyne functionalized probes for the tagging and trapping of enzyme intermediates was completed and the subsequent compatibility of such dyes with cellular incubation conditions was proven.

Type of Work:

Thesis (Doctorates > Ph.D.)

Award Type:

Doctorates > Ph.D.

Supervisor(s):