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/ Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Cell (0092-8674) 2009 imex curation Casino P., Rubio V., Marina A. amarina@ibv.csic.es 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 9 in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight by comassie molecular weight estimation by coomasie staining weight by comassie Casino P., Rubio V., Marina A. amarina@ibv.csic.es Accepted 2009-DEC-10 by JYOTI. 23-Dec-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 9 imex curation Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Cell (0092-8674) 2009 imex curation Casino P., Rubio V., Marina A. amarina@ibv.csic.es 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 9 in vitro In vitro phosphotransferase phosphotransferase assay phosphotransferase weight by comassie molecular weight estimation by coomasie staining weight by comassie Casino P., Rubio V., Marina A. Phosphorylation status of proteins/protein complex monitored by change in molecular weight following native gel electrophoresis. amarina@ibv.csic.es Accepted 2009-DEC-10 by JYOTI. 23-Dec-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 9 imex curation Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Cell (0092-8674) 2009 imex curation Casino P., Rubio V., Marina A. amarina@ibv.csic.es 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 9 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Casino P., Rubio V., Marina A. amarina@ibv.csic.es Accepted 2009-DEC-10 by JYOTI. 23-Dec-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 9 imex curation Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Structural insight into partner specificity and phosphoryl transfer in two-component signal transduction. Cell (0092-8674) 2009 imex curation Casino P., Rubio V., Marina A. amarina@ibv.csic.es 23-Dec-2009: Contacted by JYOTI. jyoti Only protein-protein interactions 9 in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Casino P., Rubio V., Marina A. Phosphorylation monitored by incorporation of 32P-labelled phosphate onto protein. amarina@ibv.csic.es Accepted 2009-DEC-10 by JYOTI. 23-Dec-2009: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 9 imex curation q9wzv7_thema TM_0853 protein protein thema Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099) MEEIKEQNNRLFLLLSIMIMEFSNAKTESDILQGLLNLVRKVVDIKEVVLVDENKRKIWGRDIDIKRFEEFIDWSIRQSNPVFVEDELGYVGIVPVVKQDRMFGSLIVLLNHQPSMEETEIFKVLSFLSAVVLENIKLYRELEETYNYVNVILNGLPEGIFVYSNGEIKFQNEKFKEENFPDEVLRKAISLSEEAISLRTQRVGEVISGEEFFSITSIPLILGSEVQALTIVENVTESKELERLKRIDRMKTEFIANISHELRTPLTAIKAYAETIYNSLGELDLSTLKEFLEVIIDQSNHLENLLNELLDFSRLERKSLQINREKVDLCDLVESAVNAIKEFASSHNVNVLFESNVPCPVEAYIDPTRIRQVLLNLLNNGVKYSKKDAPDKYVKVILDEKDGGVLIIVEDNGIGIPDHAKDRIFEQFYRVDSSLTYEVPGTGLGLAITKEIVELHGGRIWVESEVGKGSRFFVWIPKDRAGEDNRQDN DIP protein Q9WZV7 original sequence version: 94 40F19F843A4EDDB4 q9wyt9_thema TM_0468 protein protein thema Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099) MSKKVLLVDDSAVLRKIVSFNLKKEGYEVIEAENGQIALEKLSEFTPDLIVLDIMMPVMDGFTVLKKLQEKEEWKRIPVIVLTAKGGEEDESLALSLGARKVMRKPFSPSQFIEEVKHLLNE DIP protein Q9WYT9 original sequence version: 86 D946EB06B5F279E0 a0a0h3k900_staae Sensor protein kinase WalK phoR NWMN_1585 protein protein staae Staphylococcus aureus (strain Newman) MMKFHHRLMLLISSILIISFVTLGIIIHASITYVLSEKHEQALATDARSYIYLVQDNEIDKVKKIVKQQNIDLLITKHDKKVFSSGKLKAIPNSDSYIQKRDVFIFNKKVNGYHVWIKGYNNDITEMHWTLWKYLILTCLVVLICLYFASRSFKRTLIRPIQEVTYATQLLANGYYHIRVPESNVVETKALFVSTNDLARRLQKLNNEQKIQSNRLKTTIENIPSAILMIDRNGKIVVANKAYYEQFNISHNIEQVGYHGYVNTEIEQLILESFKVEKPIYEQLEVAINQVHAKYFDISCVPILTRSQKSLQGILVVMHDITNLKQLENLRREFVANVSHELKTPITSIKGFAETLIDGAKNDAESLDMFLNIILKESNRIESLVTDLLDLSHIEQHTELDTDYMNLSDLTRRIIDNMMTQANQKNISIHTDIEKDVIVKAQESKIAQVITNLLTNAINYSYEDGDINVRVYRDDFRVIFEVQDFGIGIKLEDQQRIFERFYRVDKARSRDSGGTGLGLSITKHIVEAHQGNIEVNSQVGKGSTFKVILKDYKE 4A0AA7F2CE43233D phor_bacsu Alkaline phosphatase synthesis sensor protein PhoR phoR BSU29100 protein protein bacsu Bacillus subtilis MNKYRVRLFSVFVVCMILVFCVLGLFLQQLFETSDQRKAEEHIEKEAKYLASLLDAGNLNNQANEKIIKDAGGALDVSASVIDTDGKVLYGSNGRSADSQKVQALVSGHEGILSTTDNKLYYGLSLRSEGEKTGYVLLSASEKSDGLKGELWGMLTASLCTAFIVIVYFYSSMTSRYKRSIESATNVATELSKGNYDARTYGGYIRRSDKLGHAMNSLAIDLMEMTRTQEMQRDRLLTVIENIGSGLIMIDGRGFINLVNRSYAKQFHINPNHMLRRLYHDAFEHEEVIQLVEDIFMTETKKCKLLRLPIKIERRYFEVDGVPIMGPDDEWKGIVLVFHDMTETKKLEQMRKDFVANVSHELKTPITSIKGFTETLLDGAMEDKEALSEFLSIILKESERLQSLVQDLLDLSKIEQQNFTLSIETFEPAKMLGEIETLLKHKADEKGISLHLNVPKDPQYVSGDPYRLKQVFLNLVNNALTYTPEGGSVAINVKPREKDIQIEVADSGIGIQKEEIPRIFERFYRVDKDRSRNSGGTGLGLAIVKHLIEAHEGKIDVTSELGRGTVFTVTLKRAAEKSA 0C4EE530A79A477B q9wzv7-q9wyt9-1 3 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component 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) direct interaction direct interaction R-factor: 23.6%, Free R-factor: 28.4% 2.8A One molecule of the complex formed by one HK853CP dimer and two RR468 monomers was found in the asymmetric unit of the crystals Table 1 q9wzv7-q9wyt9-2 1 6 unspecified role unspecified role neutral component neutral component Q9WYT9:p.Ile17Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain I N Wed May 02 17:27:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile17asn' Tue Jun 30 18:19:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile17Asn' Q9WYT9:p.Phe20Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F N Wed May 02 16:44:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe20asn' Tue Jun 30 18:34:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe20Asn' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9WZV7:p.Tyr272Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y D Wed May 02 19:31:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr272asp' Tue Jun 30 19:41:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr272Asp' Q9WZV7:p.Thr267Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T D Wed May 02 14:30:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr267asp' Tue Jun 30 17:46:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr267Asp' ecoli Escherichia coli (strain K12) direct interaction direct interaction R-factor: 23.6%, Free R-factor: 28.4% 2.8A One molecule of the complex formed by one HK853CP dimer and two RR468 monomers was found in the asymmetric unit of the crystals Table 1 q9wzv7-1 4 5 self self self self region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation true Visualization of 32P radioactivity after SDS-PAGE of [γ-32P]ATP-autophosphorylated HK853CP Fig. 3C, 5A-E q9wzv7-q9wyt9-3 2 5 phosphate donor phosphate donor neutral component neutral component Q9WZV7:p.Tyr272Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y D Wed May 02 15:43:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr272asp' Tue Jun 30 17:45:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr272Asp' Q9WZV7:p.Thr267Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T D Wed May 02 15:44:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr267asp' Tue Jun 30 19:40:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr267Asp' 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) 6 phosphate acceptor phosphate acceptor neutral component neutral component Q9WYT9:p.Ile17Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain I N Wed May 02 19:14:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile17asn' Tue Jun 30 16:01:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile17Asn' Q9WYT9:p.Phe20Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F N Wed May 02 19:17:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe20asn' Tue Jun 30 20:20:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe20Asn' ecoli Escherichia coli (strain K12) phosphotransfer phosphotransfer reaction phosphotransfer In agreement with these effects, each of the four mutations abolished or greatly hampered both the phosphoryl transfer from the HK to the RR (determined as RR468-triggered dephosphorylation of 32Pnot, vert, similarHK853CP) and the HK-catalyzed dephosphorylation of Pnot, vert, similarRR468 (assessed by electrophoretic quantification of the phosphorylated fraction of RR468) Fig. 3D q9wzv7-q9wyt9-4 2 5 phosphate donor phosphate donor neutral component neutral component Q9WZV7:p.Tyr272Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y D Wed May 02 15:54:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr272asp' Tue Jun 30 15:51:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr272Asp' Q9WZV7:p.Thr267Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T D Wed May 02 18:13:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr267asp' Tue Jun 30 17:14:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr267Asp' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9WZV7:p.His260Ala mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain H A Wed May 02 18:51:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his260ala' Tue Jun 30 20:15:04 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His260Ala' ecoli Escherichia coli (strain K12) 6 phosphate donor phosphate donor neutral component neutral component Q9WYT9:p.Ile17Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain I N Wed May 02 20:25:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile17asn' Tue Jun 30 17:10:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile17Asn' Q9WYT9:p.Phe20Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F N Wed May 02 19:28:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe20asn' Tue Jun 30 18:21:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe20Asn' ecoli Escherichia coli (strain K12) phosphotransfer phosphotransfer reaction phosphotransfer In agreement with these effects, each of the four mutations abolished or greatly hampered both the phosphoryl transfer from the HK to the RR (determined as RR468-triggered dephosphorylation of 32Pnot, vert, similarHK853CP) and the HK-catalyzed dephosphorylation of Pnot, vert, similarRR468 (assessed by electrophoretic quantification of the phosphorylated fraction of RR468) Fig. 3D, Supp. Fig. 7 q9wzv7-2 4 5 putative self putative self putative self putative self Q9WZV7:p.His260Ala Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain H A Wed May 02 16:51:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his260ala' Tue Jun 30 17:24:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His260Ala' c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range 5 putative self putative self putative self putative self region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range Q9WZV7:p.His260Ala Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain H A Wed May 02 17:06:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his260ala' Tue Jun 30 18:49:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His260Ala' ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Visualization of 32P radioactivity after SDS-PAGE of [γ-32P]ATP-autophosphorylated HK853CP Fig. 3C phor-1 4 7 self self self self cytoplasmic 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) phosphorylation phosphorylation reaction phosphorylation true To test whether cis phosphorylation occurs in other HKs, we analyzed the phosphorylation mechanism of PhoR from S. aureus Fig. 5F phor-2 4 8 putative self putative self putative self putative self ecoli Escherichia coli (strain K12) 8 putative self putative self putative self putative self 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) phosphorylation phosphorylation reaction phosphorylation To test whether cis phosphorylation occurs in other HKs, we analyzed the phosphorylation mechanism of PhoR from S. aureus Fig. 5F phor-3 4 7 putative self putative self putative self putative self cytoplasmic 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) 7 putative self putative self putative self putative self cytoplasmic 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 A0A0H3K900:p.Asn456Ala Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain N A Wed May 02 14:24:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn456ala' Tue Jun 30 20:21:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn456Ala' ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Time course of autophosphorylation of S. aureus PhoR heterodimers of the normal cytoplasmic portion (short PhoR) and of an elongated form (long PhoR) depending on whether the wild-type or the N456A mutant of the short form was present in the heterodimer. Fig. 5F