Design and Synthesis of New Chemical Probes and Their Applications to Understanding Protein Structure and Function by NMR

Understanding protein structure and function, including the mechanisms of protein-protein and protein-small molecule interactions, is challenging and requires the use of a range of sophisticated techniques including X-ray crystallography, NMR and cryoEM. Recently we designed a powerful in vitro NMR assay to monitor protein-bound biosynthetic transformations using a single carbon-13 tracer in a complex assay without any need for purification.

The aim of this project is to build upon this exciting foundation and expand this basic technique for wide application by the scientific community. We will focus on the elucidation of enzyme mechanisms to facilitate the development of biocatalysts for use in the clean and efficient synthesis of biological active targets (with a focus on antibiotics) and analysis of the conformation of proteins, including, for example, a-synuclein which is implicated in Parkinson's disease.

This project will involve the design and synthesis of the NMR probes with selective incorporation of carbon-13 label(s). Syntheses of isotopically labelled compounds are challenging, due to the limited availability of suitably labelled starting materials and their costs. Hence efficient synthetic strategies are required. These probes will either be ligated to the protein or used as the small molecule component of the system under study. Assessment will then be conducted by high field NMR spectroscopy. The benefits of replacing protons adjacent to the carbon-13 label with deuterium to help to sharpen the NMR signals of large systems will be investigated.