PT - JOURNAL ARTICLE AU - Joury S van ‘t Klooster AU - Tan-Yun Cheng AU - Hendrik R Sikkema AU - Aike Jeucken AU - D. Branch Moody AU - Bert Poolman TI - Periprotein membrane lipidomics and the role of lipids in transporter function in yeast AID - 10.1101/2020.01.12.903161 DP - 2020 Jan 01 TA - bioRxiv PG - 2020.01.12.903161 4099 - http://biorxiv.org/content/early/2020/01/13/2020.01.12.903161.short 4100 - http://biorxiv.org/content/early/2020/01/13/2020.01.12.903161.full AB - The yeast plasma membrane is segregated into domains: the Micro-Compartment-of-Can1 (MCC) and Pma1 (MCP) have a different protein composition, but their lipid composition is largely unknown. We extracted proteins residing in these microdomains via stoichiometric capture of lipids and proteins in styrene-maleic-acid-lipid-particles (SMALPs). We purified SMALPs by affinity chromatography and quantitatively analyzed the lipids by mass spectrometry and their role in transporter function. We found that phospholipid and sterol concentrations are similar for MCC and MCP, but sphingolipids are enriched in MCP. Ergosterol is depleted from the periprotein lipidome, whereas phosphatidylserine is enriched relative to the bulk of the plasma membrane. Phosphatidylserine, non-bilayer lipids and ergosterol are essential for activity of Lyp1; the transporter also requires a balance of saturated/unsaturated fatty acids. We propose that proteins can function in the yeast plasma membrane by the disordered state of surrounded lipids and diffuse slowly in domains of high lipid order.Impact statement Membrane protein-specific lipidomics provides information on the organization of the yeast plasma membrane and the functioning of solute transporters