Supramolecular organisation and dynamics of mannosylated phosphatidylinositol lipids in the mycobacterial plasma membrane

Tuberculosis (TB) is caused by the bacterium Mycobacterium tuberculosis (Mtb), a pathogen that claims ∼1.5 million lives annually. Drug susceptible infections are treated with a 6-month regime which is riddled with serious side-effects. Missed doses can contribute towards drug resistant and multi-drug resistant TB - an issue that is a global health concern. It is clear we need new methods to treat TB, but this pathogen is incredibly difficult to study experimentally and has physical barriers to prevent small molecule entry. The Mtb cell wall comprises of glycans and Mtb specific lipids that assemble into four discrete layers. The most well studied is the outer membrane which houses the mycolic acids but less is known about the inner layers - especially the plasma membrane. Bansal-Mutalik et al. (2014) proposed a unique composition, with over 50% of the lipids being mannosylated phosphatidylinositol lipids (phosphatidyl-myoinositol mannosides, PIMs). How PIMs contribute to the structure and function of the Mtb plasma membrane is yet to be fully elucidated. In this work we used multiscale molecular dynamics (MD) simulations to examine the structure-function relationship of the PIM lipids found in mycobacteria. The biophysical properties of the lipids in a bilayer were also investigated, looking at diffusion, rigidity, clustering, and density. The model was validated using membrane proteins and antibiotics known to interact with them, and experimental results were replicated in these simulations. Our results provide a robust workflow to assemble a mycobacterial membrane in silico to study how biophysical properties change as lipid composition changes, and hence how that might influence the passage of antibiotics into the cell.