Impact statementMembrane protein-specific lipidomics provides information on the organization of the yeast plasma membrane and the functioning of solute transporters 2 Abstract 21 22The yeast plasma membrane is segregated into domains: the Micro-Compartment-of-Can1 (MCC) 23and Pma1 (MCP) have a different protein composition, but their lipid composition is largely 24 unknown. We extracted proteins residing in these microdomains via stoichiometric capture of lipids 25and proteins in styrene-maleic-acid-lipid-particles (SMALPs). We purified SMALPs by affinity 26 chromatography and quantitatively analyzed the lipids by mass spectrometry and their role in 27 transporter function. We found that phospholipid and sterol concentrations are similar for MCC and 28MCP, but sphingolipids are enriched in MCP. Ergosterol is depleted from the periprotein lipidome, 29whereas phosphatidylserine is enriched relative to the bulk of the plasma membrane. 30Phosphatidylserine, non-bilayer lipids and ergosterol are essential for activity of Lyp1; the 31 transporter also requires a balance of saturated/unsaturated fatty acids. We propose that proteins 32can function in the yeast plasma membrane by the disordered state of surrounded lipids and diffuse 33 slowly in domains of high lipid order. 34 35