Identification of novel phospholipid binding proteins in Saccharomyces cerevisiae.
A proteomics approach was used to search for novel phospholipid binding proteins in Saccharomyces cerevisiae. Phospholipids were immobilized on a solid support and the lipids were probed with soluble yeast protein extracts. From this, the phosphatidic acid binding proteins were eluted and identified by mass spectrometry. Thirteen proteins were identified and 11 of these were previously unknown lipid binding proteins. The protein-lipid interactions identified would not have been predicted using bioinformatics approaches as none possessed a known lipid binding motif. A subset of the identified proteins was purified to homogeneity and determined to directly bind phospholipids immobilized on a solid support or organized into liposomes. This simple approach could be systematically applied to perform an exhaustive screen for soluble lipid binding proteins in S. cerevisiae or other organisms.
Fernández-Murray
J Pedro
JP
Department of Pediatrics, Atlantic Research Centre, Dalhousie University, 5849 University Avenue, Room C-302, Halifax, NS, Canada B3H 4H7.
McMaster
Christopher R
CR
D020816:Amino Acid Motifs; D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D010712:Phosphatidic Acids / Q000737:chemistry* / Q000378:metabolism; D049294:Phospholipid Transfer Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000378:metabolism
D010712:Phosphatidic Acids; D049294:Phospholipid Transfer Proteins; D029701:Saccharomyces cerevisiae Proteins