IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. The Journal of biological chemistry 2009 imex curation Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. j.c.holthuis@uu.nl 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast ub reconstruction ubiquitin reconstruction ub reconstruction predetermined predetermined participant predetermined Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. j.c.holthuis@uu.nl Accepted 2017-FEB-28 AT 11:21 GMT AT 11:21 GMT by MGT The Journal of biological chemistry 2009 Only protein-protein interactions imex curation imex curation These have a do not export stamp on as an experimental annotation (explanation in Description) so need to be in a separate expt).; Comment added Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. The Journal of biological chemistry 2009 imex curation Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. j.c.holthuis@uu.nl 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast tap tandem affinity purification TAP TAP tap western blot western blot Immuno blot Immuno blot Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. no This interaction does not occur in vivo and appears to be caused by the c-terminal myc tag on the LEM3 protein j.c.holthuis@uu.nl Accepted 2017-FEB-28 AT 11:21 GMT AT 11:21 GMT by MGT The Journal of biological chemistry 2009 Only protein-protein interactions imex curation imex curation These have a do not export stamp on as an experimental annotation (explanation in Description) so need to be in a separate expt).; Comment added Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. The Journal of biological chemistry 2009 imex curation Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. j.c.holthuis@uu.nl 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast tap tandem affinity purification TAP TAP tap western blot western blot Immuno blot Immuno blot Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. Digitonin-solubilized membranes were subjected to Ni2+-NTA chromatography, and peak fractions were pooled and passed over anti-HA antibody-coupled agarose beads. The complex was further purified using anti-myc Abs. j.c.holthuis@uu.nl Accepted 2017-FEB-28 AT 11:21 GMT AT 11:21 GMT by MGT The Journal of biological chemistry 2009 Only protein-protein interactions imex curation imex curation These have a do not export stamp on as an experimental annotation (explanation in Description) so need to be in a separate expt).; Comment added Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. The Journal of biological chemistry 2009 imex curation Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. j.c.holthuis@uu.nl 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro atpase assay atpase assay predetermined predetermined participant predetermined Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. Rate of ATPase hydrolysis was determined by a phosphomolybdate colorimetric assay The assay was performed in the presence of different phospholipids. j.c.holthuis@uu.nl Accepted 2017-FEB-28 AT 11:21 GMT AT 11:21 GMT by MGT The Journal of biological chemistry 2009 Only protein-protein interactions imex curation imex curation These have a do not export stamp on as an experimental annotation (explanation in Description) so need to be in a separate expt).; Comment added Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. Cdc50p plays a vital role in the ATPase reaction cycle of the putative aminophospholipid transporter Drs2p. The Journal of biological chemistry 2009 imex curation Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. j.c.holthuis@uu.nl 27-Mar-2018: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro phosphotransferase phosphotransferase assay phosphotransferase weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Lenoir G., Williamson P., Puts CF., Holthuis JC., Lenoir G., Williamson P., Puts CF., Holthuis JC. I've deliberately not put GO annotation on here, as the EC number corresponds to phospholipid-translocating ATPase activity but that isn't what they are measuring here. I have not included the SERCA data as that appears to be a very crude preparation. The assay was performed in the presence of different lipid mixtures (0.1 mg/ml PC/PA or PC/PA/PS) j.c.holthuis@uu.nl Accepted 2017-FEB-28 AT 11:21 GMT AT 11:21 GMT by MGT The Journal of biological chemistry 2009 Only protein-protein interactions imex curation imex curation These have a do not export stamp on as an experimental annotation (explanation in Description) so need to be in a separate expt).; Comment added Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method cdc50_yeast Cell division control protein 50 YCR94W YCR094W CDC50 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVSLFKRGKAPPLTKEGPTSKKPPNTAFRQQRLKAWQPILSPQSVLPLLIFVACIFTPIGIGLIVSATKVQDLTIDYSHCDTKASTTAFEDIPKKYIKYHFKSKVENKPQWRLTENENGEQSCELQFEIPNDIKKSIFIYYKITNFYQNHRRYVQSFDTKQILGEPIKKDDLDTSCSPIRSREDKIIYPCGLIANSMFNDTFSQVLSGIDDTEDYNLTNKHISWSIDRHRFKTTKYNASDIVPPPNWMKKYPDGYTDENLPDIHTWEEFQVWMRTAAFPKFYKLTLKNESASLPKGKYQMNIELNYPISLFGGTKSFVLTTNGAIGGRNMSLGVLYLIVAGLCALFGIIFLVKLIFQPRAMGDHTYLNFDDEENEDYEDVHAENTTLREIL 565575CA402C0C91 565575CA402C0C91 atc8_yeast Probable phospholipid-transporting ATPase DNF3 DNF3 Aminophospholipid translocase Phospholipid translocase YM8520.11C YMR162C protein protein yeast Saccharomyces cerevisiae Baker's yeast MGIADGQRRRSSSLRTQMFNKHLYDKYRGRTDDEIELEDINESKTFSGSDNNDKDDRDETSGNYAAEEDYEMEEYGSPDVSYSIITKILDTILDRRRTFHSKDGRHIPIILDHNAIEYKQAATKRDGHLIDERFNKPYCDNRITSSRYTFYSFLPRQLYAQFSKLANTYFFIVAVLQMIPGWSTTGTYTTIIPLCVFMGISMTREAWDDFRRHRLDKEENNKPVGVLVKDGNNDAQEVYTLPSSVVSSTAYLTKSAAAENNPPLNDDRNSSQGHFLDTHFNNFELLKNKYNVHIHQKKWEKLRVGDFVLLTQDDWVPADLLLLTCDGENSECFVETMALDGETNLKSKQPHPELNKLTKAASGLANINAQVTVEDPNIDLYNFEGNLELKNHRNDTIMKYPLGPDNVIYRGSILRNTQNVVGMVIFSGEETKIRMNALKNPRTKAPKLQRKINMIIVFMVFVVATISLFSYLGHVLHKKKYIDQNKAWYLFQADAGVAPTIMSFIIMYNTVIPLSLYVTMEIIKVVQSKMMEWDIDMYHAETNTPCESRTATILEELGQVSYIFSDKTGTLTDNKMIFRKFSLCGSSWLHNVDLGNSEDNFEDNRDNTNSLRLPPKAHNGSSIDVVSIGDQNVLDRLGFSDAPIEKGHRPSLDNFPKSRNSIEYKGNSSAIYTGRPSMRSLFGKDNSHLSKQASVISPSETFSENIKSSFDLIQFIQRYPTALFSQKAKFFFLSLALCHSCLPKKTHNESIGEDSIEYQSSSPDELALVTAARDLGYIVLNRNAQILTIKTFPDGFDGEAKLENYEILNYIDFNSQRKRMSVLVRMPNQPNQVLLICKGADNVIMERLHDRELAAKKMADICTSTKERKDAEAELVLQQRKSLERMVDEEAMARTSLRNSLSSVPRASLSLQAVRKSLSMKNSRTRDPEKQIDSIDQFLETVKKSDQEIGSVVNKSRKSLHKQQIEKYGPRISIDGTHFPNNNVPIDTRKEGLQHDYDTEILEHIGSDELILNEEYVIERTLQAIDEFSTEGLRTLVYAYKWIDIGQYENWNKRYHQAKTSLTDRKIKVDEAGAEIEDGLNLLGVTAIEDKLQDGVSEAIEKIRRAGIKMWMLTGDKRETAINIGYSCMLIKDYSTVVILTTTDENIISKMNAVSQEVDSGNIAHCVVVIDGATMAMFEGNPTYMSVFVELCTKTDSVICCRASPSQKALMVSNIRNTDPNLVTLAIGDGANDIAMIQSADIGVGIAGKEGLQASRVSDYSIGQFRFLLKLLFVHGRYNYIRTSKFMLCTFYKEITFYFTQLIYQRYTMFSGSSLYEPWSLSMFNTLFTSLPVLCIGMFEKDLKPMTLLTVPELYSYGRLSQGFNWLIFMEWVILATTNSLIITFLNVVMWGMSSLSDNTMYPLGLINFTAIVALINVKSQFVEMHNRNWLAFTSVVLSCGGWLVWCCALPILNNTDQIYDVAYGFYNHFGKDITFWCTSLVLALLPITLDIVYKTFKVMIWPSDSDIFAELEQKSDIRKKLELGAYSEMRQGWTWDKDPSTFTRYTDKVLSRPRTNSRASAKTHNSSIYSMSNGNVDHSSKKNFFGNSSKKSSERYEVLPSGKLIKRPSLKTQSSKDSIGGNITTKLTKKLKLPSRNVEDEDVNQIIQARLKDLE A20A823BEB401184 lem3_yeast Alkylphosphocholine resistance protein LEM3 LEM3 BRE3 Ro-sensitive 3 Brefeldin-A sensitivity protein 3 ROS3 N0333 YNL323W protein protein yeast Saccharomyces cerevisiae Baker's yeast MVNFDLGQVGEVFRRKDKGAIVSGDNPEEEEDVDASEFEEDEVKPVRTKNRRPKEDAFTQQRLAAINPVLTPRTVLPLYLLIAVVFVIVGGCILAQNSKVDEVTIYYQDCMTNATSSWSDIPSEHWQFVFHKYKTYNTAPQWRFVDDESDDFTKQRGTCQIRFTTPSDMKNNVYLNYVLEKFAANHRRYVLSFSEDQIRGEDASYETVHDATGINCKPLSKNADGKIYYPCGLIANSMFNDTFPLQLTNVGDTSNNYSLTNKGINWESDKKRYKKTKYNYTQIAPPPYWEKMYPDGYNETNIPDIQDWEEFQNWMRPGAFDKITKLIRINKNDTLPAGEYQLDIGLHWPVLEFNGKKGIYLTHGSHLGGRNPFLGIVYLIGGCICAAMALILLTFWLFGGRKIADASSLSWNMK 55BDA1F32251B4E6 55BDA1F32251B4E6 atc3_yeast Probable phospholipid-transporting ATPase DRS2 DRS2 FUN38 YAL026C protein protein yeast Saccharomyces cerevisiae Baker's yeast MNDDRETPPKRKPGEDDTLFDIDFLDDTTSHSGSRSKVTNSHANANYIPPSHVLPEETIDLDADDDNIENDVHENLFMSNNHDDQTSWNANRFDSDAYQPQSLRAVKPPGLFARFGNGLKNAFTFKRKKGPESFEMNHYNAVTNNELDDNYLDSRNKFNIKILFNRYILRKNVGDAEGNGEPRVIHINDSLANSSFGYSDNHISTTKYNFATFLPKFLFQEFSKYANLFFLCTSAIQQVPHVSPTNRYTTIGTLLVVLIVSAMKECIEDIKRANSDKELNNSTAEIFSEAHDDFVEKRWIDIRVGDIIRVKSEEPIPADTIILSSSEPEGLCYIETANLDGETNLKIKQSRVETAKFIDVKTLKNMNGKVVSEQPNSSLYTYEGTMTLNDRQIPLSPDQMILRGATLRNTAWIFGLVIFTGHETKLLRNATATPIKRTAVEKIINRQIIALFTVLIVLILISSIGNVIMSTADAKHLSYLYLEGTNKAGLFFKDFLTFWILFSNLVPISLFVTVELIKYYQAFMIGSDLDLYYEKTDTPTVVRTSSLVEELGQIEYIFSDKTGTLTRNIMEFKSCSIAGHCYIDKIPEDKTATVEDGIEVGYRKFDDLKKKLNDPSDEDSPIINDFLTLLATCHTVIPEFQSDGSIKYQAASPDEGALVQGGADLGYKFIIRKPNSVTVLLEETGEEKEYQLLNICEFNSTRKRMSAIFRFPDGSIKLFCKGADTVILERLDDEANQYVEATMRHLEDYASEGLRTLCLAMRDISEGEYEEWNSIYNEAATTLDNRAEKLDEAANLIEKNLILIGATAIEDKLQDGVPETIHTLQEAGIKIWVLTGDRQETAINIGMSCRLLSEDMNLLIINEETRDDTERNLLEKINALNEHQLSTHDMNTLALVIDGKSLGFALEPELEDYLLTVAKLCKAVICCRVSPLQKALVVKMVKRKSSSLLLAIGDGANDVSMIQAAHVGVGISGMEGMQAARSADIAVGQFKFLKKLLLVHGSWSYQRISVAILYSFYKNTALYMTQFWYVFANAFSGQSIMESWTMSFYNLFFTVWPPFVIGVFDQFVSSRLLERYPQLYKLGQKGQFFSVYIFWGWIINGFFHSAIVFIGTILIYRYGFALNMHGELADHWSWGVTVYTTSVIIVLGKAALVTNQWTKFTLIAIPGSLLFWLIFFPIYASIFPHANISREYYGVVKHTYGSGVFWLTLIVLPIFALVRDFLWKYYKRMYEPETYHVIQEMQKYNISDSRPHVQQFQNAIRKVRQVQRMKKQRGFAFSQAEEGGQEKIVRMYDTTQKRGKYGELQDASANPFNDNNGLGSNDFESAEPFIENPFADGNQNSNRFSSSRDDISFDI 5745B92901F8E1AE 5745B92901F8E1AE atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) atc5_yeast Phospholipid-transporting ATPase DNF1 DNF1 YER166W SYGP-ORF7 Flippase DNF1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSGTFHGDGHAPMSPFEDTFQFEDNSSNEDTHIAPTHFDDGATSNKYSRPQVSFNDETPKNKREDAEEFTFNDDTEYDNHSFQPTPKLNNGSGTFDDVELDNDSGEPHTNYDGMKRFRMGTKRNKKGNPIMGRSKTLKWARKNIPNPFEDFTKDDIDPGAINRAQELRTVYYNMPLPKDMIDEEGNPIMQYPRNKIRTTKYTPLTFLPKNILFQFHNFANVYFLVLIILGAFQIFGVTNPGLSAVPLVVIVIITAIKDAIEDSRRTVLDLEVNNTKTHILEGVENENVSTDNISLWRRFKKANSRLLFKFIQYCKEHLTEEGKKKRMQRKRHELRVQKTVGTSGPRSSLDSIDSYRVSADYGRPSLDYDNLEQGAGEANIVDRSLPPRTDCKFAKNYWKGVKVGDIVRIHNNDEIPADIILLSTSDTDGACYVETKNLDGETNLKVRQSLKCTNTIRTSKDIARTKFWIESEGPHSNLYTYQGNMKWRNLADGEIRNEPITINNVLLRGCTLRNTKWAMGVVMFTGGDTKIMLNSGITPTKKSRISRELNFSVVINFVLLFILCFVSGIANGVYYDKKGRSRFSYEFGTIAGSAATNGFVSFWVAVILYQSLVPISLYISVEIIKTAQAAFIYGDVLLYNAKLDYPCTPKSWNISDDLGQVEYIFSDKTGTLTQNVMEFKKCTINGVSYGRAYTEALAGLRKRQGIDVETEGRREKAEIAKDRDTMIDELRALSGNSQFYPEEVTFVSKEFVRDLKGASGEVQQRCCEHFMLALALCHSVLVEANPDNPKKLDLKAQSPDEAALVATARDVGFSFVGKTKKGLIIEMQGIQKEFEILNILEFNSSRKRMSCIVKIPGLNPGDEPRALLICKGADSIIYSRLSRQSGSNSEAILEKTALHLEQYATEGLRTLCIAQRELSWSEYEKWNEKYDIAAASLANREDELEVVADSIERELILLGGTAIEDRLQDGVPDCIELLAEAGIKLWVLTGDKVETAINIGFSCNLLNNEMELLVIKTTGDDVKEFGSEPSEIVDALLSKYLKEYFNLTGSEEEIFEAKKDHEFPKGNYAIVIDGDALKLALYGEDIRRKFLLLCKNCRAVLCCRVSPSQKAAVVKLVKDSLDVMTLAIGDGSNDVAMIQSADVGIGIAGEEGRQAVMCSDYAIGQFRYLARLVLVHGRWSYKRLAEMIPEFFYKNMIFALALFWYGIYNDFDGSYLYEYTYMMFYNLAFTSLPVIFLGILDQDVNDTISLVVPQLYRVGILRKEWNQRKFLWYMLDGLYQSIICFFFPYLVYHKNMIVTSNGLGLDHRYFVGVYVTTIAVISCNTYVLLHQYRWDWFSGLFIALSCLVVFAWTGIWSSAIASREFFKAAARIYGAPSFWAVFFVAVLFCLLPRFTYDSFQKFFYPTDVEIVREMWQHGHFDHYPPGYDPTDPNRPKVTKAGQHGEKIIEGIALSDNLGGSNYSRDSVVTEEIPMTFMHGEDGSPSGYQKQETWMTSPKETQDLLQSPQFQQAQTFGRGPSTNVRSSLDRTREQMIATNQLDNRYSVERARTSLDLPGVTNAASLIGTQQNN 3CC3FBA8ADDE8960 3CC3FBA8ADDE8960 yn8s_yeast Probable phospholipid translocase non-catalytic subunit CRF1 CDC50/ROS3 family protein 1 N3453 YNR048W protein protein yeast Saccharomyces cerevisiae Baker's yeast MGLILRWKEKKQLSSKQNAQKSRKPANTSFRQQRLKAWQPILSPQSVLPLLILMACVFAPIGIGLVVSTISVQRLVVNYTECDALAPAKHFETIPSEYVDYHFSKKVAVQPQWMVLTDPELGNQTCRIQFEVPNHIKKSTYVYYRLTNFNQNYREYVQSLDLDQLKGKALIGNDLDPNCDPLRTVENKTIFPCGLIANSMFNDTFGTTLTGVNDTADYLLTTKGIAWDTDSHRYGKTEYNASDIVPPPNWAKLFPNGYTDDNIPDLQNWEQFKIWMRTAALPNFYKLAMKNETNGLGKGIYIADIELNYPVRSFYGTKSFVLTTNSIIGAGNEALGIVYLIVAGIATLFAILFLIKVIFKPRPMHDHSYLNFENSDTPFDESSVVSIPLREIL 9BCA199AA3D23969 drs2-cdc50-2 3 7 unspecified role unspecified role prey prey c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 10 unspecified role unspecified role bait bait P39524:p.Asp560Asn mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain D N Wed May 02 16:10:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp560asn' Tue Jun 30 19:55:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp560Asn' n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figs. 2, 4A, 5C, 6A drs2-lem3-1 2 9 unspecified role unspecified role prey prey c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 10 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Fig. 5C cdc50-drs2-1 3 7 unspecified role unspecified role bait bait c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 10 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range physical association physical association Figs 4A, 5D drs2_cdc50_yeast-1 5 35 putative self putative self putative self putative self DRS2 n-terminal tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range CDC50 c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast phosphotransfer phosphotransfer reaction phosphotransfer true N-ethylmaleimide (CHEBI:44485) - an inhibitor of the aminophospholipid translocase activity Orthovanadate (CHEBI:46442 ) GO term deliberately not added as I can't make a match to what is shown in the expt. Figs3A-E, 4B, 5E The aspartate at position 560 (Asp-560) in Drs2p is part of the DKTGTLT signature sequence and is modified to form the aspartyl-phosphate intermediate. All of the VO4-sensitive phosphorylation was abolished in a asp560asn mutant. This indicates that wild type DRS2 co-purified with CDC50 undergoes VO4-sensitive phosphorylation from ATP at Asp-560. dnf1-lem3-3 1 9 unspecified role unspecified role prey prey n-terminal nub nub n-term range n-terminal range n-term range n-term range n-terminal range n-term range 12 unspecified role unspecified role bait bait c-terminal cub cub c-term range C-terminal range c-term range c-term range C-terminal range c-term range P32660:p.Asp667Asn mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain D N Wed May 02 16:59:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp667asn' Tue Jun 30 16:51:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp667Asn' P32660:p.Asp667Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain D E Wed May 02 16:46:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp667glu' Tue Jun 30 19:11:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp667Glu' physical association physical association Figs. 6 B/E dnf3-crf1-1 1 13 unspecified role unspecified role prey prey n-terminal nub nub n-term range n-terminal range n-term range n-term range n-terminal range n-term range 8 unspecified role unspecified role bait bait c-terminal cub cub c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figs. 6B drs2-cdc50-1 1 7 unspecified role unspecified role prey prey n-terminal nub nub n-term range n-terminal range n-term range n-term range n-terminal range n-term range 10 unspecified role unspecified role bait bait P39524:p.Glu342Gln mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E Q Wed May 02 20:13:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu342gln' Tue Jun 30 20:15:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu342Gln' P39524:p.Gly341Leu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain G L Wed May 02 20:29:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly341leu' Tue Jun 30 20:06:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly341Leu' P39524:p.Asp560Asn mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain D N Wed May 02 17:01:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp560asn' Tue Jun 30 17:52:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp560Asn' P39524:p.Asp560Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain D E Wed May 02 18:30:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp560glu' Tue Jun 30 16:43:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp560Glu' c-terminal cub cub c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figs. 6B-D, 7B-D drs2_cdc50_yeast-2 4 11 enzyme target enzyme target substrate substrate unspecified role unspecified role 35 enzyme enzyme neutral component neutral component DRS2 n-terminal tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range CDC50 c-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast atpase reaction atpase reaction Orthovanadate (CHEBI:46442 ) Supp Fig 2 drs2-cdc50_yeast DRS2-CDC50 P4-ATPase complex CDC5:DRS2 Aminophospholipid translocase complex APLT complex DRS2-CDC50 phospholipid flippase complex 7 unspecified role unspecified role drs2 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 enzyme enzyme cdc50 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 65 67 direct interaction direct interaction yeast Saccharomyces cerevisiae Baker's yeast stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction A membrane pump which actively translocates, or flips, phospholipids across cell membranes from the exoplasmic to the cytoplasmic leaflet of the lipid bilayer. This generates and maintains membrane lipid asymmetry, a property essential for a wide variety of cellular processes such as vesicle budding and intracellular vesicle trafficking. Transport is accomplished by cyclic changes between two main enzyme conformations, during which the DRS2 ATPase is phosphorylated by ATP at a conserved aspartate residue (Asp-560) and subsequently dephosphorylated. These processes are coupled to vectorial transport and counter-transport by a controlled opening and closing of cytoplasmic and exoplasmic pathways, which give access to the ion-binding sites that are buried inside the membrane-spanning region of the pump The membrane domain of DRS2 serves as the pathway for translocation of lipid substrates across cell membranes and is predicted to contain 10 transmembrane segments. CDC50 is a membrane glycoprotein and appears to contact DRS2 at multiple sites to promote correct protein folding, exit of the P4-ATPase complex from the endoplasmic reticulum, and formation of a functionally active flippase complex. Accepted 2017-DEC-20 by LIVIAP