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Ab GTPase Ras GTPase Rab GTPase Ras GTPase Rho GTPase Rho GTPase Rab GTPase Rab GTPase Rab GTPase Putative little GTPase activator Rab GTPase Rho GTPase Rab GTPase Rab GTPase Ras GTPase Rab GTPase Rab GTPase Ras GTPase Rab GTPase Rab GTPaseB, C, D D B, D DDD C, D DD CBD D14385444 545224 196alg2 pdi1 rplPPutative alpha-1,3-mannosyltransferase Protein disulfide isomerase 60S ribosomal acidic ribosomal protein PA, C A, B A(Continued on following web page)ec.asm.orgEukaryotic CellLipid Droplets in DictyosteliumTABLE 1 (Continued)MASCOT score by conditionb 1st 66 63 2nd 60 77 53 77 48 67 3rd 205 245 160 158 98 Mean MASCOT scorec 111 103 79 71 69 53 50 40 Presence in LDs of other cell sort(s)d B, D B, C, D A A, C A, C B A, CProtein group and identification no.a HDAC8 Inhibitor Compound DDB0231975 DDB0233663 DDB0233863 DDB0231366 DDB0231241 DDB0231191 DDB0191150 DDB0214854 Cytoskeleton components DDB0191168 DDB0191134 DDB0185047 DDB0185015 DDB0219923 DDB0305338 Miscellaneous DDB0266658 DDB0191175 DDB0184006 DDB0238661 DDB0190526 DDB0304659 DDBa bGene name ugt1 rtnlC alg11 rpl4 rpl10 alg1 rplGene product Putative glycosyltransferase Heat shock protein 70 with ER retention signal Reticulon family protein Putative alpha-1,CXCR Antagonist site 2-mannosyltransferase 60S ribosomal protein L4 S60 ribosomal protein L10 Putative glycosyltransferase S60 ribosomal protein L129 61 101245 43 275 92 20679 77 82 133 65742 836 487356 319 282 225 91hspB efaA11 hspE-1 act comAHeat shock protein 70 cytoplasmic, F-actin connected Elongation aspect 1 , F-actin binding and bundling Heat shock protein 70, cytoplasmic, F-actin linked Actin Comitin, F-actin-binding Golgi protein Heat shock protein 70, cytoplasmicA, B, D C A, B B, C A, B, D151 36 124 108 96 59366 78 68 109 43 671,181 1,089 726 261566 401 306 160 104 42aifC cadA ldpA netDPutative apoptosis inducing aspect Cell adhesion molecule, plasma membrane and cytosol Lipid droplet membrane protein NET4 Putative transmembrane metallo-phosphoesterase Includes protein phosphatase 2C-related domain No recognizable featuresB BProteins are grouped as outlined by function. Rows in bold highlight proteins investigated within this work. Identification numbers are from dictyBase. MASCOT scores of the MS/MS ion search procedure correspond for the frequency of peptide identification from 3 independent preparations as specified in the text. c Solely to provide a measure for sorting proteins within the groups the imply MASCOT score was determined. d Letters A to D indicate irrespective of whether the protein was also identified as a constituent of lipid droplets in yeast (38), enterocytes (39), muscle cells (40), and/or pancreatic beta cells (41), respectively.ate circumstances, we transfected a plasmid expressing the human homologue tagged at its C-terminal finish with GFP (24) into two mammalian cell lines and obtained a clear association with LD540-labeled structures induced by oleate feeding (Fig. 5E and F).DISCUSSIONFlow of fatty acids. The natural meals sources of Dictyostelium, i.e., several species of bacteria, contain fatty acids mainly in esterified type. Endosomal lipases are believed to liberate fatty acids, which can partition within the membrane vectorially. The direction of this pathway is brought about by the activity from the FcsA enzyme on the cytoplasmic face of your endosome, which adds a coenzyme A moiety to produce the activated fatty acid, which is then further funneled into metabolism (15). Within the laboratory, it truly is a lot more reproducible to induce the formation of lipid droplets that stor.

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