Dynamic S-adenosyl-L-methionine cofactor analogues for use in the methyltransferase-directed labelling of DNA

Wilkinson, Andrew (2020). Dynamic S-adenosyl-L-methionine cofactor analogues for use in the methyltransferase-directed labelling of DNA. University of Birmingham. Ph.D.

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Current bioconjugation strategies are a versatile tool for the introduction of non-natural functionality to biomolecules including DNA, RNA, proteins. However, current strategies lack the versatility required to undergo sequential functionalisations. This is an area of increasing interest in chemical biology, as it facilitates multiple analyses of an individual biomolecule to help develop a wider understanding of its role within a system. Methyltransferase directed bioconjugation offers a site-selective approach for the introduction of a variety of functional groups to biomolecules, in complex mixtures.

Here we present the design and synthesis of a series of ‘doubly-activated’ methyltransferase cofactor analogues, based on the structure of the naturally occurring cofactor S-adenosyl-L-methionine. Specifically, we developed two cofactor analogues which included a dynamic covalent moiety and could undergo sequential modifications once introduced to a biomolecule of interest. Our approach uses a centrally located Schiff base in the form of either an oxime or hydrazone, with terminal azide functionality to allow for further modification once introduced to a biomolecule of interest.

Through a series of proof of concept experiments we have been able to rigorously test each cofactor analogue developed in the transalkylation of DNA with methyltransferases. We have then gone on to show that when present within the DNA backbone, a hydrazone moiety provides an excellent platform for the repeated functionalisation of DNA. This work is now being developed further in attempts to utilise this chemistry for unique bioconjugation applications in epigenetics and microscopy.

Type of Work: Thesis (Doctorates > Ph.D.)
Award Type: Doctorates > Ph.D.
Neely, Robertr.k.neely@bham.ac.ukUNSPECIFIED
Fernandez-Trillo, Franciscof.fernandez-trillo@bham.ac.ukUNSPECIFIED
Licence: All rights reserved
College/Faculty: Colleges (2008 onwards) > College of Engineering & Physical Sciences
School or Department: School of Chemistry
Funders: Engineering and Physical Sciences Research Council
Subjects: Q Science > QD Chemistry
URI: http://etheses.bham.ac.uk/id/eprint/10095


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