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/ Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae. Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae. The Journal of biological chemistry 2002 imex curation Lobo S., Greentree WK., Linder ME., Deschenes RJ. Robert-deschenes@uiowa.edu Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast pull down pull down anti tag western anti tag western blot anti tag western Lobo S., Greentree WK., Linder ME., Deschenes RJ. Robert-deschenes@uiowa.edu Accepted 2017-FEB-06 AT 08:50 GMT AT 08:50 GMT by MGT The Journal of biological chemistry 2002 imex curation Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae. Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae. The Journal of biological chemistry 2002 imex curation Lobo S., Greentree WK., Linder ME., Deschenes RJ. Robert-deschenes@uiowa.edu Only protein-protein interactions in vitro In vitro palmitoylase assay palmitoylase assay palmitoylase assay tag visualisation tag visualisation Lobo S., Greentree WK., Linder ME., Deschenes RJ. Included as the 'partial purification' looks pretty impressive in Fig. 2 Robert-deschenes@uiowa.edu Accepted 2017-FEB-06 AT 08:50 GMT AT 08:50 GMT by MGT The Journal of biological chemistry 2002 imex curation Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae. Identification of a Ras palmitoyltransferase in Saccharomyces cerevisiae. The Journal of biological chemistry 2002 imex curation Lobo S., Greentree WK., Linder ME., Deschenes RJ. Robert-deschenes@uiowa.edu Only protein-protein interactions ecoli Escherichia coli (strain K12) pull down pull down anti tag western anti tag western blot anti tag western Lobo S., Greentree WK., Linder ME., Deschenes RJ. Robert-deschenes@uiowa.edu Accepted 2017-FEB-06 AT 08:50 GMT AT 08:50 GMT by MGT The Journal of biological chemistry 2002 imex curation 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 erfb_yeast Palmitoyltransferase ERF2 DHHC cysteine-rich domain-containing protein ERF2 Ras protein acyltransferase ERF2 YLR246W protein protein yeast Saccharomyces cerevisiae Baker's yeast MALVSRRSTRSESTSITKEEHTGEGSLTKLFFRWLVTLEGDQDINDGKGYISLPNVSNYIFFLGGRFRTVKGAKPLWLGVLLAIVCPMVLFSIFEAHKLWHTQNGYKVLVIFFYYFWVITLASFIRTATSDPGVLPRNIHLSQLRNNYQIPQEYYNLITLPTHSSISKDITIKYCPSCRIWRPPRSSHCSTCNVCVMVHDHHCIWVNNCIGKRNYRFFLIFLLGAILSSVILLTNCAIHIARESGGPRDCPVAILLLCYAGLTLWYPAILFTYHIFMAGNQQTTREFLKGIGSKKNPVFHRVVKEENIYNKGSFLKNMGHLMLEPRGPSFVSARKPHEAGDWRFMDLSPAHSFEKIQKI B1FF42A2370F064B ras2_yeast Ras-like protein 2 RAS2 ASC1 CTN5 GLC5 YNL098C N2198 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPLNKSNIREYKLVVVGGGGVGKSALTIQLTQSHFVDEYDPTIEDSYRKQVVIDDEVSILDILDTAGQEEYSAMREQYMRNGEGFLLVYSITSKSSLDELMTYYQQILRVKDTDYVPIVVVGNKSDLENEKQVSYQDGLNMAKQMNAPFLETSAKQAINVEEAFYTLARLVRDEGGKYNKTLTENDNSKQTSQDTKGSGANSVPRNSGGHRKMSNAANGKNVNSSTTVVNARNASIESKTGLAGNQATNGKTQTDRTNIDNSTGQAGQANAQSANTVNNRVNNNSKAGQVSNAKQARKQQAAPGGNTSEASKSGSGGCCIIS 6CAAAC531EEAE92F 6CAAAC531EEAE92F palmitoleoyl-coa palmitoleoyl-CoA (Z)-9-hexadecenoyl-CoA cis-9-hexadecenoyl-CoA cis-9-hexadecenoyl-coenzyme A palmitoleoyl-coenzyme A palmitoleyl-CoA palmitoleyl-coenzyme A 3'-phosphoadenosine 5'-{3-[(3R)-3-hydroxy-2,2-dimethyl-4-({3-[(2-{[(9Z)-hexadec-9-enoyl]sulfanyl}ethyl)amino]-3-oxopropyl}amino)-4-oxobutyl] dihydrogen diphosphate} small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) stearoyl-coa stearoyl-CoA C18:0-CoA C18:0-coenzyme A octadecanoyl-CoA octadecanoyl-coenzyme A S-stearoyl-CoA S-stearoylcoenzyme A Stearoyl-CoA stearoyl-coenzyme A Stearyl coenzyme A Stearyl-CoA 3'-phosphoadenosine 5'-(3-{(3R)-3-hydroxy-2,2-dimethyl-4-[(3-{[2-(octadecanoylsulfanyl)ethyl]amino}-3-oxopropyl)amino]-4-oxobutyl} dihydrogen diphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) palmitoyl-coa palmitoyl-CoA small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) oleoyl-coa oleoyl-CoA (9Z)-octadec-9-enoyl-CoA (9Z)-octadec-9-enoyl-coenzyme A (9Z)-Octadecenoyl-CoA cis-9-octadecenoyl-CoA cis-9-octadecenoyl-coenzyme A cis-Delta(9)-octadecenoyl-CoA cis-Delta(9)-octadecenoyl-coenzyme A Oleoyl-CoA Oleoyl-coenzyme A S-[(9Z)-octadec-9-enoyl]-CoA S-[(9Z)-octadec-9-enoyl]-coenzyme A S-oleoyl-CoA S-Oleoylcoenzyme A 3'-phosphoadenosine 5'-{3-[(3R)-3-hydroxy-2,2-dimethyl-4-({3-[(2-{[(9Z)-octadec-9-enoyl]sulfanyl}ethyl)amino]-3-oxopropyl}amino)-4-oxobutyl] dihydrogen diphosphate} small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) erfd_yeast Ras modification protein ERF4 YOL110W HRC237 SHR5 ERF4 Hyperactive Ras suppressor protein 5 protein protein yeast Saccharomyces cerevisiae Baker's yeast MCDSHQKEEDNANTSERALFFNYHEFSYSFYEDLGSEDAKPTEHDEDHKLCITHFPNVYAARGSAEFQVTRVVRVPRRFDESRSSLETPQFSTQLPGSEPAAIVGDDGTSFVRCGRYDIGDHVFGCSSVSPLSEYLSAAELAEVVHRVNGFLLREEGEVFGWRNLSGLLLDMLTGGLWSWVLGPLLSRPVFQESLALEQYVAQLNSPGGLLHERGVRLVLPRRSGCLSLDFVVPRPK 28F66E12B491ADAD shr5-erf2-2 1 5 unspecified role unspecified role prey prey n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range Q06551:p.Cys189Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:38:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys189ser' Tue Jun 30 19:06:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys189Ser' Q06551:p.His201Ala 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 H A Wed May 02 15:12:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his201ala' Tue Jun 30 16:14:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His201Ala' 11 unspecified role unspecified role bait bait n-terminal 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 physical association physical association Fig. 1B, 2 erf2_shr5_1-palmitoyl_coa-1 2 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 23 enzyme enzyme neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 6 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast palmitoylation palmitoylation reaction palmitoylation Fig. 1C, Fig. 4 Erf2 Cys189Ser and His201Ala mutants abolished the catalytic activity of the enzyme complex shr5-erf2-1 3 5 unspecified role unspecified role prey prey n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 11 unspecified role unspecified role bait bait n-terminal 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 physical association physical association Fig. 3C erf2_shr5_1-palmitoyl_coa-2 2 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 23 enzyme enzyme neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) palmitoylation palmitoylation reaction palmitoylation Fig. 3B Erf2 Cys189Ser and His201Ala mtants abolished the catalytic activity of the enzyme complex erf2_shr5_1-palmitoyl_coa-3 2 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 23 enzyme enzyme neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 6 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P01120:p.Cys318Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain C A Wed May 02 16:34:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys318ala' Tue Jun 30 18:23:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys318Ala' yeast Saccharomyces cerevisiae Baker's yeast palmitoylation palmitoylation reaction palmitoylation Table 1 PAT activity 2720 pmol palmitate transferred/min/μmol substrate erf2_shr5_1-palmitoyl_coa-4 2 23 putative self putative self neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined putative self interaction putative self interaction Fig. 4 Formation of a [3H]palmitoylated Erf2p acyl-enzyme intermediate. Erf2 Cys189Ser and His201Ala mutants abolished the catalytic activity of the enzyme complex erf2_shr5_1-palmitoyl_coa-5 2 9 competitor competitor competitor neutral component neutral component 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 23 enzyme enzyme neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 6 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P01120:p.Cys318Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain C A Wed May 02 18:10:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys318ala' Tue Jun 30 16:52:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys318Ala' yeast Saccharomyces cerevisiae Baker's yeast palmitoylation palmitoylation reaction palmitoylation Fig 5 erf2_shr5_1-palmitoleoyl_coa-1 2 7 competitor competitor competitor neutral component neutral component 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 23 enzyme enzyme neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 6 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P01120:p.Cys318Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain C A Wed May 02 16:21:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys318ala' Tue Jun 30 18:40:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys318Ala' yeast Saccharomyces cerevisiae Baker's yeast palmitoylation palmitoylation reaction palmitoylation Fig 5 erf2_shr5_1-palmitoyl_coa-6 2 8 competitor competitor competitor neutral component neutral component 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 23 enzyme enzyme neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 6 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P01120:p.Cys318Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain C A Wed May 02 14:30:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys318ala' Tue Jun 30 18:51:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys318Ala' yeast Saccharomyces cerevisiae Baker's yeast palmitoylation palmitoylation reaction palmitoylation Fig 5 erf2_shr5_1-oleoyl_coa-1 2 10 competitor competitor competitor neutral component neutral component 9 enzyme target enzyme target substrate substrate neutral component neutral component region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 23 enzyme enzyme neutral component neutral component shr5 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 erf2 flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range yeast Saccharomyces cerevisiae Baker's yeast 6 enzyme target enzyme target substrate substrate neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P01120:p.Cys318Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain C A Wed May 02 15:23:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys318ala' Tue Jun 30 18:01:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys318Ala' yeast Saccharomyces cerevisiae Baker's yeast palmitoylation palmitoylation reaction palmitoylation Fig 5 erf2-shr5-1 Palmitoyltransferase ERF2/SHR5 complex ERF2:SHR5 Erf2/Erf4 PAT complex Palmitoyltransferase ERF2-SHR5 complex Erf2-Erf4 PAT complex 5 enzyme enzyme shr5 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role erf2 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 102 104 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 Palmitoyltransferase complex that catalyzes the palmitoylation of proteins through a two step reaction - the autopalmitoylation of the enzyme to create a palmitoyl-ERF2 intermediate followed by the transfer of the palmitoyl moiety to the RAS substrate. Palmitoyl-CoA serves as the palmitate donor. ERF2 appears to have a direct role in palmitate transfer based on its ability to form an acyl-intermediate. Accepted 2017-FEB-23 by BMELDAL