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/ Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Scientific reports 2014 imex curation Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za Accepted 2016-MAY-26 AT 14:29 BST AT 14:29 BST by MGT Scientific reports 2014 Only protein-protein interactions imex curation imex curation Unfortunately, I don't think this has been down a gel filtration column if you read the text "The expressed proteins were subsequently purified using Ni2+-based IMAC. SDS-PAGE analysis (Fig. 3a) ..". 2 baits are allowed if there is no other option, and I can't think of one in this case. PD domains come from PMID:22124281, Ref. 1 in Supp materials cited in Supp. Table 2; I can't see a way round the enzyme problem - if you look at Fig. 1 the catalytic site is formed cooperatively between the two proteins. All else done, Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Scientific reports 2014 imex curation Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by staining molecular weight estimation by staining weight by staining Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. The complex was first purified by Ni Chromatography, then subjected to SEC (Superdex 200). estrauss@sun.ac.za Accepted 2016-MAY-26 AT 14:29 BST AT 14:29 BST by MGT Scientific reports 2014 Only protein-protein interactions imex curation imex curation Unfortunately, I don't think this has been down a gel filtration column if you read the text "The expressed proteins were subsequently purified using Ni2+-based IMAC. SDS-PAGE analysis (Fig. 3a) ..". 2 baits are allowed if there is no other option, and I can't think of one in this case. PD domains come from PMID:22124281, Ref. 1 in Supp materials cited in Supp. Table 2; I can't see a way round the enzyme problem - if you look at Fig. 1 the catalytic site is formed cooperatively between the two proteins. All else done, Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Scientific reports 2014 imex curation Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro decarboxxylation assay decarboxylation assay decarboxxylation assay predetermined predetermined participant predetermined Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. Phosphopantothenoylcysteine decarboxylase (PPCDC) assay couples the formation of the 4′-phosphopantetheine product to E. coli phosphopantetheine adenylyltransferase (EcPPAT), the next enzyme in the CoA pathway. The assay links the pyrophosphate released by this enzyme to the oxidation of NADH, which is followed spectrophotometrically. estrauss@sun.ac.za Accepted 2016-MAY-26 AT 14:29 BST AT 14:29 BST by MGT Scientific reports 2014 Only protein-protein interactions imex curation imex curation Unfortunately, I don't think this has been down a gel filtration column if you read the text "The expressed proteins were subsequently purified using Ni2+-based IMAC. SDS-PAGE analysis (Fig. 3a) ..". 2 baits are allowed if there is no other option, and I can't think of one in this case. PD domains come from PMID:22124281, Ref. 1 in Supp materials cited in Supp. Table 2; I can't see a way round the enzyme problem - if you look at Fig. 1 the catalytic site is formed cooperatively between the two proteins. All else done, 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 Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Scientific reports 2014 imex curation Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro crosslink cross-linking study crosslink weight by staining molecular weight estimation by staining weight by staining Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. Cross-linking agent - Disuccinimidyl suberate estrauss@sun.ac.za Accepted 2016-MAY-26 AT 14:29 BST AT 14:29 BST by MGT Scientific reports 2014 Only protein-protein interactions imex curation imex curation Unfortunately, I don't think this has been down a gel filtration column if you read the text "The expressed proteins were subsequently purified using Ni2+-based IMAC. SDS-PAGE analysis (Fig. 3a) ..". 2 baits are allowed if there is no other option, and I can't think of one in this case. PD domains come from PMID:22124281, Ref. 1 in Supp materials cited in Supp. Table 2; I can't see a way round the enzyme problem - if you look at Fig. 1 the catalytic site is formed cooperatively between the two proteins. All else done, Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Scientific reports 2014 imex curation Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) pull down pull down weight by staining molecular weight estimation by staining weight by staining Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za Accepted 2016-MAY-26 AT 14:29 BST AT 14:29 BST by MGT Scientific reports 2014 Only protein-protein interactions imex curation imex curation Unfortunately, I don't think this has been down a gel filtration column if you read the text "The expressed proteins were subsequently purified using Ni2+-based IMAC. SDS-PAGE analysis (Fig. 3a) ..". 2 baits are allowed if there is no other option, and I can't think of one in this case. PD domains come from PMID:22124281, Ref. 1 in Supp materials cited in Supp. Table 2; I can't see a way round the enzyme problem - if you look at Fig. 1 the catalytic site is formed cooperatively between the two proteins. All else done, Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Complex stability and dynamic subunit interchange modulates the disparate activities of the yeast moonlighting proteins Hal3 and Vhs3. Scientific reports 2014 imex curation Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za 22-Jun-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Abrie JA., Molero C., Ariño J., Strauss E. I haven't added the CD as I don't think its indicating binding, also the mutants in Fig 7 aren't shown against a control so I've ignored those. estrauss@sun.ac.za Accepted 2016-MAY-26 AT 14:29 BST AT 14:29 BST by MGT Scientific reports 2014 Only protein-protein interactions imex curation imex curation Unfortunately, I don't think this has been down a gel filtration column if you read the text "The expressed proteins were subsequently purified using Ni2+-based IMAC. SDS-PAGE analysis (Fig. 3a) ..". 2 baits are allowed if there is no other option, and I can't think of one in this case. PD domains come from PMID:22124281, Ref. 1 in Supp materials cited in Supp. Table 2; I can't see a way round the enzyme problem - if you look at Fig. 1 the catalytic site is formed cooperatively between the two proteins. All else done, cab3_yeast Coenzyme A biosynthesis protein 3 YKL088W CAB3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MTDEKVNSDQNMNGKQGVNLISSLPTTQVPVSILTNKERRKSIHDESNFERSDSHEDQSKSNSNRRNIYKNDYSTNLRDFSFANLKQNSERNKDGHEIQINTSMPANTNGQQKRFSPSLPSAVSFTVPEVERLPYHRYSISNKPGKQQQQQEQLQQNQQQEEQQKAQLQEQNQRAKQQEEVKQIQEQVQKKQTERQQLIDEKERIANAIFKENTTNDGTDIRKHSVSSGTSNSEDEVDSPSMEKNSIVHMPGDFIYFNPKSNASKPITAKAAPLSANNSTHKNKEVITAPTGPRVPFTEFFQKEDDKKFHILIGATGSVATIKVPLIIDKLFKIYGPEKISIQLIVTKPAEHFLKGLKMSTHVKIWREEDAWVFDAVNKNDTSLSLNLILHHELRKWADIFLIAPLSANTLAKLANGICNNLLTSVMRDWSPLTPVLIAPAMNTFMYINPMTKKHLTSLVQDYPFIQVLKPVEKVLICGDIGMGGMREWTDIVEIVRRRINEIRKARDEETGDKEQEQEEQEGADNEDDDDEDDEEDEEDEEEEEALNETASDESNDEEDEEDEEDVKTEV 9C674C2394EFCEAB ppz1_yeast Serine/threonine-protein phosphatase PP-Z1 PPZ1 YM9571.02C YML016C protein protein yeast Saccharomyces cerevisiae Baker's yeast MGNSSSKSSKKDSHSNSSSRNPRPQVSRTETSHSVKSAKSNKSSRSRRSLPSSSTTNTNSNVPDPSTPSKPNLEVNHQRHSSHTNRYHFPSSSHSHSNSQNELLTTPSSSSTKRPSTSRRSSYNTKAAADLPPSMIQMEPKSPILKTNNSSTHVSKHKSSYSSTYYENALTDDDNDDKDNDISHTKRFSRSSNSRPSSIRSGSVSRRKSDVTHEEPNNGSYSSNNQENYLVQALTRSNSHASSLHSRKSSFGSDGNTAYSTPLNSPGLSKLTDHSGEYFTSNSTSSLNHHSSRDIYPSKHISNDDDIENSSQLSNIHASMENVNDKNNNITDSKKDPNEEFNDIMQSSGNKNAPKKFKKPIDIDETIQKLLDAGYAAKRTKNVCLKNNEILQICIKAREIFLSQPSLLELSPPVKIVGDVHGQYGDLLRLFTKCGFPPSSNYLFLGDYVDRGKQSLETILLLFCYKIKYPENFFLLRGNHECANVTRVYGFYDECKRRCNIKIWKTFIDTFNTLPLAAIVAGKIFCVHGGLSPVLNSMDEIRHVVRPTDVPDFGLINDLLWSDPTDSPNEWEDNERGVSYCYNKVAINKFLNKFGFDLVCRAHMVVEDGYEFFNDRSLVTVFSAPNYCGEFDNWGAVMSVSEGLLCSFELLDPLDSAALKQVMKKGRQERKLANQQQQMMETSITNDNESQQ 8B7F009521F72C15 8B7F009521F72C15 n-[(r)-4-phosphopantothenoyl]-l-cysteine N-[(R)-4-phosphopantothenoyl]-L-cysteine (R)-4'-Phosphopantothenoyl-L-cysteine N-((R)-4'-phosphopantothenoyl)-L-cysteine N-[(R)-4'-Phosphopantothenoyl]-L-cysteine N-{N-[(2R)-2-hydroxy-3,3-dimethyl-4-phosphonooxybutanoyl]-beta-alanyl}-L-cysteine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) vhs3_yeast Phosphopantothenoylcysteine decarboxylase subunit VHS3 Viable in a HAL3 SIT4 background protein 3 VHS3 YOR054C YOR29-05 protein protein yeast Saccharomyces cerevisiae Baker's yeast MTNKSSLKNNRKGVASNTLSGAEQANIGSSAMPDTNSTGPFSSVSSLDTPVVRKSTSPTGSQTKSIMNASGTSGAVVSNTPEPGLKRIPTVTFSDPKLGSLRSDVEQTPPNQVARQSSEKKATSVHIAAEGANQGRNLKDINTKVPKDGEASASSFSTPTSILSNADMGNNISSLLAKKLSFTGGTDSILNSDNSSDSPRKEHPHFYVEDPLHTPSVRSRSNSTSPRPSVVVNTFNPINIEREGSISKTGEPTLLESVLEEAMSPNAVSNPLKRENIMTNMDPRLPQDDGKLHVLFGATGSLSVFKLKHMIRKLEEIYGRDKICIQVILTNSATKFFAMKYMRKNKKQHNSIDTSFNSTNSNAGNITGNKKKVASLEKFSIQKTSSNSAASQTNNKQEEEKQMASTTGFPSTLGGSRTYSNSSNVVSQHPQIELPAHIQFWTDQDEWDVWRQRTDPVLHIELRRWADILVVAPLTANTLAKIALGLCDNLLTSVIRAWNPTFPIFLAPSMGSGTFNSIMTKKHFRIIQEEMPWVTVFKPSEKVMGINGDIGLSGMMDANEIVGKIVVKLGGYPDVSAGKEEEEDEDNDEEDDNKKNDTGGKDEDNDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDDEDDEDEDEDDEGKKKEDKGGLQRS A97623473CB6F605 A97623473CB6F605 coac_human Phosphopantothenoylcysteine decarboxylase CoaC PPCDC COAC MDS018 UNQ9365/PRO34154 protein protein human Homo sapiens Human MEPKASCPAAAPLMERKFHVLVGVTGSVAALKLPLLVSKLLDIPGLEVAVVTTERAKHFYSPQDIPVTLYSDADEWEIWKSRSDPVLHIDLRRWADLLLVAPLDANTLGKVASGICDNLLTCVMRAWDRSKPLLFCPAMNTAMWEHPITAQQVDQLKAFGYVEIPCVAKKLVCGDEGLGAMAEVGTIVDKVKEVLFQHSGFQQS 9E59683480032894 sis2_yeast Phosphopantothenoylcysteine decarboxylase subunit SIS2 SIS2 HAL3 YKR072C Halotolerance protein HAL3 Sit4 suppressor 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MTAVASTSGKQDADHNQSIECPRFSRGQKEILLDHEDAKGKDSIINSPVSGRQSISPTLSNATTTTTKSIMNATGTSGAVVSNTPEPGLKRVPAVTFSDLKQQQKQDSLTQLKNDSERTKSPNSNPAPVSNSIPGNHAVIPNHTNTSRTTQLSGSPLVNEMKDYDPKKKDSALKIVDTMKPDKIMATSTPISRENNKVTAKAPTSITLRKEDAQDQANNVSGQINVRSTPEETPVKQSVIPSIIPKRENSKNLDPRLPQDDGKLHVLFGATGSLSVFKIKPMIKKLEEIYGRDRISIQVILTQSATQFFEQRYTKKIIKSSEKLNKMSQYESTPATPVTPTPGQCNMAQVVELPPHIQLWTDQDEWDAWKQRTDPVLHIELRRWADILVVAPLTANTLSKIALGLCDNLLTSVIRAWNPSYPILLAPSMVSSTFNSMMTKKQLQTIKEEMSWVTVFKPSEKVMDINGDIGLGGMMDWNEIVNKIVMKLGGYPKNNEEEDDDEDEEEDDDEEEDTEDKNENNNDDDDDDDDDDDDDDDDDDDDDDDDEDEDEAETPGIIDKHQ 19A9A475145DA7AB ppz1-sis2-2 7 9 unspecified role unspecified role bait bait c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association Fig 2A The E. coli DnaK chaperone copurifies with the PPZ1 bait as a contaminant. ppz1-sis2-3 2 9 unspecified role unspecified role neutral component neutral component c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association Fig 2D cab3-sis2-1 6 8 unspecified role unspecified role prey prey pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 11 unspecified role unspecified role bait bait pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 13 unspecified role unspecified role bait bait pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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. 3A sis2-cab3-1 6 8 unspecified role unspecified role prey prey pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role bait bait pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 35 37 direct interaction direct interaction Fig. 3A vhs3-cab3-1 6 8 unspecified role unspecified role prey prey pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 11 unspecified role unspecified role bait bait pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 41 43 direct interaction direct interaction Fig. 3A sis2-cab3-4 3 10 enzyme target enzyme target substrate substrate neutral component neutral component 48 enzyme enzyme neutral component neutral component P36076 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P36024 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q08438 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P36024 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 Q08438 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 ecoli Escherichia coli (strain K12) decarboxylation decarboxylation reaction decarboxylation Fig. 3B sis2-cab3-3 3 8 enzyme enzyme neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 13 enzyme enzyme neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 10 enzyme target enzyme target substrate substrate neutral component neutral component 58 56 decarboxylation decarboxylation reaction decarboxylation Fig. 3B vhs3-cab3-2 3 8 enzyme enzyme neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 ancillary ancillary neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 10 enzyme target enzyme target substrate substrate neutral component neutral component decarboxylation decarboxylation reaction decarboxylation Fig. 3B n_[(r)_4_phosphopantothenoyl]_l_cysteine-ppcdc-1 3 10 enzyme target enzyme target substrate substrate neutral component neutral component 12 enzyme enzyme neutral component neutral component decarboxylation decarboxylation reaction decarboxylation Fig. 3B sis2-1 5 13 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 direct interaction direct interaction Fig. 4A sis2-2 5 13 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 direct interaction direct interaction Fig. 4A The presence of dimers is assumed to be due to incomplete crosslinking. vhs3-3 5 11 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 direct interaction direct interaction Fig. 4A vhs3-1 5 11 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 direct interaction direct interaction Fig. 4A The presence of dimers is assumed to be due to incomplete crosslinking. ppcdc-1 1 12 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Fig. 4B sis2-5 1 13 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction Fig. 4C Additional higher molecular weight oligomers appear to form. vhs3-4 1 11 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction Fig. 4D sis2-cab3-2 1 8 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 99 97 direct interaction direct interaction Fig. 4E sis2-cab3-5 1 8 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 11 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 13 unspecified role unspecified role neutral component neutral component pd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 4F ppcdc_yeast Phosphopantothenoylcysteine decarboxylase complex PPCDC complex CAB3:SIS2:VHS3 PPC-DC complex CoaC complex CoA caroxylase complex Hal3-Vhs3-Cab3 complex 8 unspecified role unspecified role sis2 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain vhs3 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 11 enzyme enzyme cab3 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 13 enzyme enzyme cab3 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 108 113 111 109 physical association physical association 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 Catalyses the third step of the coenzyme A (CoA) biosynthetic pathway, namely the decarboxylation of 4'-phosphopantothenoylcysteine to form 4′-phosphopantetheine. May potentially be part of a larger Coenzyme A-synthesizing protein complex (CPX-396) but this is not yet clear. The synthesis of CoA involves the phosphorylation of pantothenate (vitamin B5) to 4'-phosphopantothenate, to which a cysteine is then added to form 4'-phospho-N-pantothenoylcysteine (PCC). PPC is decarboxylated to 4'-phosphopantetheine by phosphopantothenoylcysteine decarboxylase (this complex). 4'-phosphopantetheine is adenylylated to form dephospho-CoA which is then phosphorylated to form coenzyme A. Two catalytically essential residues are required - His that is required for the first step of the enzyme’s two-step mechanism (an oxidative decarboxylation), and a Cys that is necessary for the second step (the reduction of the reaction intermediate). These are found on opposite sides of the active site, with VHS3 contributing the His residue whilst CAB3 contributes the Cys residue to give a complex with a single functional active site. Heterotrimer Accepted 2016-APR-27 by BMELDAL