A chemical biology approach to interrogate the zDHHC enzymes

At the core of pharmaceutical research is the discovery and validation of new targets for drug discovery. This project resides in the Healthcare Technologies theme, aligning with the research areas of synthetic organic chemistry, computational chemistry and the growth area of chemical biology to deliver chemical tools to interrogate zDHHC enzymes as targets for discovery research.
S-Acylation, the attachment of fatty acids onto cysteine residues, regulates the activity of a wide range of proteins and is catalysed by the zDHHC family of enzymes. There are no chemical tools available to understand the biology of these enzymes. Based upon recent joint work between the supervisors we will use an emerging technology in chemical biology to deliver small molecule probes for this class of enzyme.
The work will be performed in three distinct phases:
Phase 1: The preparation, isolation and purification of zDHHC enzymes. This will be performed in the Chamberlain laboratory using established techniques. Initially three distinct targets will be prepared: hDHHC2 catalytic domain. hDHHC17 catalytic domain. hDHHC17 ankyrin repeat domain.
Phase 2: This will be performed with project partners GSK. Using a recently prepared library of photoaffinity fragments our enzymes will be screened against this 550 member library. Using an established screening protocol protein will be incubated with individual photoaffinity fragments then irradiated at 302 nm. Analysis using intact protein mass spectrometry will identify any binding fragments. Follow up assays will confirm hit identification.
Phase 3: Fragment hits identified will be elaborated using a structure guided iterative medicinal chemistry campaign to improve activity. In-house primary screens will guide this work. The goal of this phase of the work is to deliver a sub-micromolar inhibitor for each of the targets examined. Subsequent work if time permits will look for selectivity between the hDHHC2 catalytic domain and the hDHHC17 catalytic domain.
Outcome: It is expected this work will deliver the first inhibitors of the zDHHC family of enzymes that will enable investigation of the fundamental biology of these proteins.