Insight into the GAC biosynthesis machinery and how GAC is linked to Peptidoglycan in S. pyogenes cells.

点击翻译按钮获取中文摘要

Are GAC proteins forming a multi-protein complex (MPC)?I will employ bacterial two-hybrid and bacterial three-hybrid assays to further investigate interactions of GAC proteins. Microscopy techniques will be utilised to analyse colocalisation of each protein. To understand the stoichiometry of each GAC protein, isotope labelling and cross-linking mass spectrometry will be employed. This data will allow us, for the first time, to build a MPC model. Providing a starting point to elucidate the detailed arrangement of each GAC protein within the MPC (cryogenic electron microscopy).How is the mature GAC transferred from the lipid onto peptidoglycan?I aim to employ bioinformatics to identify potential functional and transmembrane domains of the LytR-Cps2A-Psr (LCP) family. Microscopy will be utilised to visualise the location of LCP proteins in relation to the cell membrane. Molecular cloning will be used to perform deletion assays, radiolabelling assays and Western blot analysis on nickel chromatography pull-down products.Project provides training in Interdisciplinary Skills