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/ The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. Cell (0092-8674) 2009 imex curation Good M., Tang G., Singleton J., Remenyi A., Lim WA. Activated kinases phosphorylate a MAPK-specific peptide LVEPLTPSGEAPNQK which is bound to fluorescence quencher. the phosphorylated peptide then binds to Gallium on the sensor bead and quenches the fluorescent polymers on the bead. lim@cmp.ucsf.edu 01-Sep-2009: Contacted by JYOTI. davet Only protein-protein interactions 5 in vitro In vitro protein kinase assay protein kinase assay western blot western blot Immuno blot Immuno blot Good M., Tang G., Singleton J., Remenyi A., Lim WA. Activated kinases phosphorylate a MAPK-specific peptide LVEPLTPSGEAPNQK which is bound to fluorescence quencher. the phosphorylated peptide then binds to Gallium on the sensor bead and quenches the fluorescent polymers on the bead. lim@cmp.ucsf.edu Accepted 2009-JUL-2 by DAVET. 01-Sep-2009: Contacted by JYOTI. Antibodies are specific for phsophorylated Fus3 and Kss1 residues. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. Cell (0092-8674) 2009 imex curation Good M., Tang G., Singleton J., Remenyi A., Lim WA. Activated kinases phosphorylate a MAPK-specific peptide LVEPLTPSGEAPNQK which is bound to fluorescence quencher. the phosphorylated peptide then binds to Gallium on the sensor bead and quenches the fluorescent polymers on the bead. lim@cmp.ucsf.edu 01-Sep-2009: Contacted by JYOTI. davet Only protein-protein interactions 5 in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Good M., Tang G., Singleton J., Remenyi A., Lim WA. Activated kinases phosphorylate a MAPK-specific peptide LVEPLTPSGEAPNQK which is bound to fluorescence quencher. the phosphorylated peptide then binds to Gallium on the sensor bead and quenches the fluorescent polymers on the bead. lim@cmp.ucsf.edu Accepted 2009-JUL-2 by DAVET. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. The Ste5 scaffold directs mating signaling by catalytically unlocking the Fus3 MAP kinase for activation. Cell (0092-8674) 2009 imex curation Good M., Tang G., Singleton J., Remenyi A., Lim WA. Activated kinases phosphorylate a MAPK-specific peptide LVEPLTPSGEAPNQK which is bound to fluorescence quencher. the phosphorylated peptide then binds to Gallium on the sensor bead and quenches the fluorescent polymers on the bead. lim@cmp.ucsf.edu 01-Sep-2009: Contacted by JYOTI. davet Only protein-protein interactions 5 in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Good M., Tang G., Singleton J., Remenyi A., Lim WA. Activated kinases phosphorylate a MAPK-specific peptide LVEPLTPSGEAPNQK which is bound to fluorescence quencher. the phosphorylated peptide then binds to Gallium on the sensor bead and quenches the fluorescent polymers on the bead. lim@cmp.ucsf.edu Accepted 2009-JUL-2 by DAVET. 01-Sep-2009: Contacted by JYOTI. Cell (0092-8674) 2009 davet Only protein-protein interactions 5 imex curation kss1_yeast Mitogen-activated protein kinase KSS1 KSS1 Kinase suppressor of SST2 YGR040W protein protein yeast Saccharomyces cerevisiae Baker's yeast MARTITFDIPSQYKLVDLIGEGAYGTVCSAIHKPSGIKVAIKKIQPFSKKLFVTRTIREIKLLRYFHEHENIISILDKVRPVSIDKLNAVYLVEELMETDLQKVINNQNSGFSTLSDDHVQYFTYQILRALKSIHSAQVIHRDIKPSNLLLNSNCDLKVCDFGLARCLASSSDSRETLVGFMTEYVATRWYRAPEIMLTFQEYTTAMDIWSCGCILAEMVSGKPLFPGRDYHHQLWLILEVLGTPSFEDFNQIKSKRAKEYIANLPMRPPLPWETVWSKTDLNPDMIDLLDKMLQFNPDKRISAAEALRHPYLAMYHDPSDEPEYPPLNLDDEFWKLDNKIMRPEEEEEVPIEMLKDMLYDELMKTME 5FA42161B5FD51B5 5FA42161B5FD51B5 ste5_yeast Protein STE5 STE5 NUL3 YDR103W YD8557.12 protein protein yeast Saccharomyces cerevisiae Baker's yeast MMETPTDNIVSPFHNFGSSTQYSGTLSRTPNQIIELEKPSTLSPLSRGKKWTEKLARFQRSSAKKKRFSPSPISSSTFSFSPKSRVTSSNSSGNEDGNLMNTPSTVSTDYLPQHPHRTSSLPRPNSNLFHASNSNLSRANEPPRAENLSDNIPPKVAPFGYPIQRTSIKKSFLNASCTLCDEPISNRRKGEKIIELACGHLSHQECLIISFGTTSKADVRALFPFCTKCKKDTNKAVQCIPENDELKDILISDFLIHKIPDSELSITPQSRFPPYSPLLPPFGLSYTPVERQTIYSQAPSLNPNLILAAPPKERNQIPQKKSNYTFLHSPLGHRRIPSGANSILADTSVALSANDSISAVSNSVRAKDDETKTTLPLLRSYFIQILLNNFQEELQDWRIDGDYGLLRLVDKLMISKDGQRYIQCWCFLFEDAFVIAEVDNDVDVLEIRLKNLEVFTPIANLRMTTLEASVLKCTLNKQHCADLSDLYIVQNINSDESTTVQKWISGILNQDFVFNEDNITSTLPILPIIKNFSKDVGNGRHETSTFLGLINPNKVVEVGNVHDNDTVIIRRGFTLNSGECSRQSTVDSIQSVLTTISSILSLKREKPDNLAIILQIDFTKLKEEDSLIVVYNSLKALTIKFARLQFCFVDRNNYVLDYGSVLHKIDSLDSISNLKSKSSSTQFSPIWLKNTLYPENIHEHLGIVAVSNSNMEAKKSILFQDYRCFTSFGRRRPNELKIKVGYLNVDYSDKIDELVEASSWTFVLETLCYSFGLSFDEHDDDDEEDNDDSTDNELDNSSGSLSDAESTTTIHIDSPFDNENATANMVNDRNLLTEGEHSNIENLETVASSVQPALIPNIRFSLHSEEEGTNENENENDMPVLLLSDMDKGIDGITRRSSFSSLIESGNNNCPLHMDYI 0435BDA0196B2D6F 0435BDA0196B2D6F ste7_yeast Serine/threonine-protein kinase STE7 STE7 D1525 YDL159W protein protein yeast Saccharomyces cerevisiae Baker's yeast MFQRKTLQRRNLKGLNLNLHPDVGNNGQLQEKTETHQGQSRIEGHVMSNINAIQNNSNLFLRRGIKKKLTLDAFGDDQAISKPNTVVIQQPQNEPVLVLSSLSQSPCVSSSSSLSTPCIIDAYSNNFGLSPSSTNSTPSTIQGLSNIATPVENEHSISLPPLEESLSPAAADLKDTLSGTSNGNYIQLQDLVQLGKIGAGNSGTVVKALHVPDSKIVAKKTIPVEQNNSTIINQLVRELSIVKNVKPHENIITFYGAYYNQHINNEIIILMEYSDCGSLDKILSVYKRFVQRGTVSSKKTWFNELTISKIAYGVLNGLDHLYRQYKIIHRDIKPSNVLINSKGQIKLCDFGVSKKLINSIADTFVGTSTYMSPERIQGNVYSIKGDVWSLGLMIIELVTGEFPLGGHNDTPDGILDLLQRIVNEPSPRLPKDRIYSKEMTDFVNRCCIKNERERSSIHELLHHDLIMKYVSPSKDDKFRHWCRKIKSKIKEDKRIKREALDRAKLEKKQSERSTH C8A75899CFBE8BDE C8A75899CFBE8BDE fus3_yeast Mitogen-activated protein kinase FUS3 FUS3 DAC2 YBL016W YBL0303 YBL03.21 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPKRIVYNISSDFQLKSLLGEGAYGVVCSATHKPTGEIVAIKKIEPFDKPLFALRTLREIKILKHFKHENIITIFNIQRPDSFENFNEVYIIQELMQTDLHRVISTQMLSDDHIQYFIYQTLRAVKVLHGSNVIHRDLKPSNLLINSNCDLKVCDFGLARIIDESAADNSEPTGQQSGMTEYVATRWYRAPEVMLTSAKYSRAMDVWSCGCILAELFLRRPIFPGRDYRHQLLLIFGIIGTPHSDNDLRCIESPRAREYIKSLPMYPAAPLEKMFPRVNPKGIDLLQRMLVFDPAKRITAKEALEHPYLQTYHDPNDEPEGEPIPPSFFEFDHYKEALTTKDLKKLIWNEIFS 5117980D20A1E7E2 5117980D20A1E7E2 ste7-kss1-7 2 6 enzyme enzyme neutral component neutral component 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 P06784:p.[Ser359Glu;Thr363Glu] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain T E Wed May 02 15:26:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser359glu,thr363glu' Tue Jun 30 19:21:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser359Glu;Thr363Glu]' 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 4 enzyme target enzyme target substrate substrate neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation We found that Ste7EE rapidly activated Kss1, Phosphorylation, and activation, of KSS1 Figs. 2B, 4B, Supp. Fig. 2B Michaelis-Menten plots showed Ste7 phosphorylation of Kss1 does not require the presence of Ste5, and had no effect of the km or Kcat of the reaction. ste7-fus3-8 2 5 unspecified role unspecified role ancillary ancillary region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P32917:p.[Asn710Ala;Glu712Ala;Lys714Ala] Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain N A certain certain sequence position certain certain certain sequence position certain E A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:48:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn710ala,glu712ala,lys714ala' Tue Jun 30 18:19:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn710Ala;Glu712Ala;Lys714Ala]' P32917:p.[Val745Ala;Tyr747_Ser748delinsAlaAla] Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain V A certain certain sequence position certain certain certain sequence position certain YS AA Thu Jul 07 12:17:18 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was val745tyr747ser748al Fri Jul 22 10:56:46 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'val745ala,tyr747ala,ser748ala' Wed May 02 19:44:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val745ala,tyr_ser747-748ala_ala' Tue Jun 30 18:34:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val745Ala;Tyr747_Ser748delinsAlaAla]' P32917:p.[Ile628Ala;Asn632Ala;Lys635Ala] Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain N A certain certain sequence position certain certain certain sequence position certain I A certain certain sequence position certain certain certain sequence position certain K A Fri Jul 22 10:55:30 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'asn632ala,ile628ala,lys635ala' Wed May 02 15:17:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile628ala,asn632ala,lys635ala' Tue Jun 30 19:36:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile628Ala;Asn632Ala;Lys635Ala]' undefined mutation Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:57:13 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'lys750ile751asp752' P32917:p.[Leu754Ala;Glu756Ala] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain L A certain certain sequence position certain certain certain sequence position certain E A Fri Jul 22 10:49:19 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys754ala,glu756ala' Wed May 02 18:36:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu754ala,glu756ala' Tue Jun 30 20:13:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu754Ala;Glu756Ala]' P32917:p.[Asn744Ala;Asp746Ala] Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain D A certain certain sequence position certain certain certain sequence position certain N A Fri Jul 22 10:59:44 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'asp746ala,asn744ala' Wed May 02 18:17:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn744ala,asp746ala' Tue Jun 30 18:23:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn744Ala;Asp746Ala]' P32917:p.[Glu765Ala;Tyr769Ala] Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y A certain certain sequence position certain certain certain sequence position certain E A Fri Jul 22 11:09:30 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr769ala,glu765ala' Wed May 02 17:23:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu765ala,tyr769ala' Tue Jun 30 18:40:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu765Ala;Tyr769Ala]' region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 7 enzyme target enzyme target substrate substrate neutral component neutral component 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) 6 enzyme enzyme neutral component neutral component 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 P06784:p.[Ser359Glu;Thr363Glu] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain T E Wed May 02 16:57:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser359glu,thr363glu' Tue Jun 30 16:11:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser359Glu;Thr363Glu]' 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation Fus3 cannot be activated by Ste7EE, unless N-Ste5 is added. Phosphorylation, and activation, of Fus3 Fig. 2B, Supp. Fig. 2B Mutation of the two N-terminal MAPK docking sites of Ste7 inhibit this interaction but no details of these mutations are given. Defective Ste5 mutants are defined as falling below the limit of detection of the assay. ste7-ste5-1 3 6 enzyme enzyme neutral component neutral component 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 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 5 unspecified role unspecified role neutral component neutral component region Fluorescein tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 In fluorescence polarization binding studies, we found that the Ste5-ms domain binds tightly to Ste7 (KD = 75nM) Fig. 3F ste7-fus3-10 1 7 enzyme target enzyme target substrate substrate neutral component neutral component 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 P16892:p.Lys42Arg Catalytically inactive mutation mutation certain certain sequence position certain certain certain sequence position certain K R Wed May 02 17:09:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys42arg' Tue Jun 30 18:53:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys42Arg' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role ancillary ancillary P32917:p.[Asn744Ala;Asp746Ala] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain N A certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:53:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn744ala,asp746ala' Tue Jun 30 18:55:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn744Ala;Asp746Ala]' P32917:p.[Val745Ala;Tyr747_Ser748delinsAlaAla] Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain YS AA certain certain sequence position certain certain certain sequence position certain V A Thu Jul 07 12:07:22 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was val745tyr747ser748al Fri Jul 22 11:08:49 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'val745ala,tyr747ala,ser748ala' Wed May 02 19:48:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val745ala,tyr_ser747-748ala_ala' Tue Jun 30 18:34:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val745Ala;Tyr747_Ser748delinsAlaAla]' undefined mutation Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:57:25 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'lys750ile751asp752' P32917:p.[Glu765Ala;Tyr769Ala] Alters kinetics of interaction mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain E A certain certain sequence position certain certain certain sequence position certain Y A Wed May 02 20:21:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu765ala,tyr769ala' Tue Jun 30 17:11:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu765Ala;Tyr769Ala]' region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain P32917:p.[Leu754Ala;Glu756Ala] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain E A certain certain sequence position certain certain certain sequence position certain L A Fri Jul 22 11:14:58 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'lys754ala,glu756ala' Wed May 02 15:15:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu754ala,glu756ala' Tue Jun 30 18:53:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu754Ala;Glu756Ala]' ecoli Escherichia coli (strain K12) 6 enzyme enzyme neutral component neutral component 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 P06784:p.[Ser359Glu;Thr363Glu] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain T E Wed May 02 14:59:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser359glu,thr363glu' Tue Jun 30 16:57:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser359Glu;Thr363Glu]' 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation Fus3 cannot be activated by Ste7EE, unless N-Ste5 is added. Phosphorylation, and activation, of Fus3 Fig. 4B, 4C, Supp. Figs. 1A, 6B, 7 Ste7EE-ND2 (which contains only one MAPK docking motif, KD 100nM), and Fus3-K42R (which is catalytically dead) were used to simplify the analyses. Kinase reactions contain 50nM Ste7EE-ND2, and a saturating concentration (1000nM, where appropriate) of truncated Ste5. The Ste5 domain enhances the kcat of Fus3 phosphorylation by Ste7EE by 5000-fold, with little effect on the Km ste7-kss1-8 1 4 enzyme target enzyme target substrate substrate neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 enzyme enzyme neutral component neutral component 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 P06784:p.[Ser359Glu;Thr363Glu] Alters kinetics of interaction mutation mutation certain certain sequence position certain certain certain sequence position certain S E certain certain sequence position certain certain certain sequence position certain T E Wed May 02 15:51:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser359glu,thr363glu' Tue Jun 30 19:05:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ser359Glu;Thr363Glu]' 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection phosphorylation phosphorylation reaction phosphorylation Ste7EE is able to phosphorylate Kss1 Phosphorylation, and activation, of KSS1 Figs. 4B, Supp. Fig. 2A Michaelis-Menten plots showed Ste7 phosphorylation of Kss1 does not require the presence of Ste5, and had no effect of the km or Kcat of the reaction.