Creating a New Photoprotein (for Healthcare Applications) using Organic Chemistry and Protein Engineering

The PhD project will involve the modification of an existing protein scaffold using site directed mutagenesis to allow it to accommodate a range of modified 'luciferins' which the student will also synthesize. The protein-luciferin combinations (photoproteins) produced will then be assessed for their ability to produce light (luminesce) in the presence of a range of activating agents including reactive oxygen species (ROS) such as singlet oxygen. Efficient photoproteins will then be evaluated in a number of applications including biosensors for the detection of ROS, and as components of protein based photodynamic therapy systems for applications in cancer. During the PhD, training will be provided in molecular biology, protein expression and engineering, as well as the characterization of the modified protein scaffolds using circular dichroism, mass spectrometry and dynamic scanning calorimetry. The synthesis strand of the project will provide training in modern multistep preparative organic chemistry and the use of high field NMR, mass spectrometry and other methods for the characterization of the new luciferins and photoprotein bioconjugates. In the final stages of the project the PhD student will be trained in mammalian cell culture and how to measure cell viability and luminescence efficiency in order to evaluate the applications of the new photoproteins. This is a multidisciplinary project that would best suit applicants with a strong interest in applying preparative chemistry to functionalising proteins and the characterisation of the resulting conjugates.