Dr. Matt Proctor, Biosciences, Sheffield
Transformation of sunlight into chemical energy during photosynthesis relies upon a network of membrane protein-protein interactions occurring within specialised lipid membranes called thylakoids. Traditionally, thylakoid proteins are isolated for study by detergent extraction, however limitations of this include the potential for perturbation of enzyme activity and protein interactions, necessitating costly and time-consuming screens to find an appropriate detergent. This project aims to identify novel photosynthetic protein complexes by avoiding disruptive detergents, furthering our understanding of energy flow and regulation during photosynthesis. We will achieve this by deploying Salipro® technology, utilising the lipid-binding protein SapA. This project allows the student to participate in all aspects of the scientific process from experimental design through to data analysis. The student will produce and purify SapA from E. coli, gaining skills in molecular biology techniques, and then following preparation of plant thylakoid membranes, will develop a method for isolating photosynthetic proteins in SapA nanodiscs. Analysis will investigate novel protein complexes by sucrose gradients, spectroscopy, immunoblotting, mass spectrometry and microscopy in comparison to classical detergent extracts. Students will be expected to present their findings orally at a research day in York in September 2023.