Effect of phophatidylcholine and phosphatidylethanolamine enrichment on the structure and function of yeast membrane.
The phospholipid composition of yeast plasma membrane was manipulated by two different methods: (i) by using two auxotrophic strains KA101 (cho1) and MC13 (Cho+) which required phospholipid bases for growth and (ii) by supplementing Saccharomyces cerevisiae (3059) cells with high concentration of choline or ethanolamine. It was possible to enrich the plasma membrane with phosphatidylcholine (PC) or phosphatidylethanolamine (PE) by both methods. The uptake of amino acids, e.g., glycine, glutamic acid, leucine, lysine methionine, phenylalanine, proline and serine, was significantly reduced in PC- or PE-enriched cells. However, the extent of reduction in transport was variable among different strains. A fluorescent probe, 1-anilino-8-naphthalene sulfonate (ANS), was used to monitor the structural changes induced by altered phospholipid composition. It was observed that the relative fluorescence intensity of bound ANS was decreased as a consequence of PC or PE enrichment. The decrease in fluorescence was probably associated with reduced number of available binding sites (n) and increased apparent dissociation constant (Kd). Furthermore, our results also suggest that a critical level of PE or PC is required for proper functioning of yeast membrane.
D000596:Amino Acids / Q000378:metabolism; D000817:Anilino Naphthalenesulfonates; D001692:Biological Transport / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000502:physiology*; D005456:Fluorescent Dyes; D007700:Kinetics; D008563:Membrane Lipids / Q000302:isolation & purification; D010713:Phosphatidylcholines / Q000494:pharmacology*; D010714:Phosphatidylethanolamines / Q000494:pharmacology*; D010743:Phospholipids / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000502:physiology*; D013045:Species Specificity; D013050:Spectrometry, Fluorescence
D000596:Amino Acids; D000817:Anilino Naphthalenesulfonates; D005456:Fluorescent Dyes; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010743:Phospholipids; C027132:1-anilino-8-naphthalenesulfonate