UniProt UniProt http://www.uniprot.org Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Journal of molecular biology 2007 imex curation Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. marmor@wistar.org 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. marmor@wistar.org Accepted 2018-JUL-04 AT 13:04 BST AT 13:04 BST by PPORRAS Journal of molecular biology 2007 Only protein-protein interactions imex curation imex curation Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Journal of molecular biology 2007 imex curation Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. marmor@wistar.org 09-May-2019: 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 Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. marmor@wistar.org Accepted 2018-JUL-04 AT 13:04 BST AT 13:04 BST by PPORRAS Journal of molecular biology 2007 Only protein-protein interactions imex curation imex curation Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Journal of molecular biology 2007 imex curation Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. marmor@wistar.org 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. marmor@wistar.org Accepted 2018-JUL-04 AT 13:04 BST AT 13:04 BST by PPORRAS Journal of molecular biology 2007 Only protein-protein interactions imex curation imex curation Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Molecular basis for bre5 cofactor recognition by the ubp3 deubiquitylating enzyme. Journal of molecular biology 2007 imex curation Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. marmor@wistar.org 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Li K., Ossareh-Nazari B., Liu X., Dargemont C., Marmorstein R. I have assumed the tags were not cleaved off for the ITC (which may well be wrong so delete if you don't agree with me) marmor@wistar.org Accepted 2018-JUL-04 AT 13:04 BST AT 13:04 BST by PPORRAS Journal of molecular biology 2007 Only protein-protein interactions imex curation imex curation bre5_yeast UBP3-associated protein BRE5 BRE5 YNR051C N3465 Brefeldin-A sensitivity protein 5 protein protein yeast Saccharomyces cerevisiae Baker's yeast MGVTVQDICFAFLQNYYERMRTDPSKLAYFYASTAELTHTNYQSKSTNEKDDVLPTVKVTGRENINKFFSRNDAKVRSLKLKLDTIDFQYTGHLHKSILIMATGEMFWTGTPVYKFCQTFILLPSSNGSTFDITNDIIRFISNSFKPYVLTDASLSQSNEENSVSAVEEDKIRHESGVEKEKEKEKSPEISKPKAKKETVKDTTAPTESSTQEKPIVDHSQPRAIPVTKESKIHTETVPSSTKGNHKQDEVSTEELGNVTKLNEKSHKAEKKAAPIKTKEGSVEAINAVNNSSLPNGKEVSDEKPVPGGVKEAETEIKPIEPQVSDAKESGNNASTPSSSPEPVANPPKMTWASKLMNENSDRISKNNTTVEYIRPETLPKKPTERKFEMGNRRDNASANSKNKKKPVFSTVNKDGFYPIYIRGTNGLREEKLRSALEKEFGKVMRITAADNFAVVDFETQKSQIDALEKKKKSIDGIEVCLERKTVKKPTSNNPPGIFTNGTRSHRKQPLKRKD 462A7D4309A69192 ubp3_yeast Ubiquitin carboxyl-terminal hydrolase 3 UBP3 Ubiquitin-specific-processing protease 3 Deubiquitinating enzyme 3 Ubiquitin thioesterase 3 YER151C protein protein yeast Saccharomyces cerevisiae Baker's yeast MNMQDANKEESYSMYPKTSSPPPPTPTNMQIPIYQAPLQMYGYTQAPYLYPTQIPAYSFNMVNQNQPIYHQSGSPHHLPPQNNINGGSTTNNNNINKKKWHSNGITNNNGSSGNQGANSSGSGMSYNKSHTYHHNYSNNHIPMMASPNSGSNAGMKKQTNSSNGNGSSATSPSYSSYNSSSQYDLYKFDVTKLKNLKENSSNLIQLPLFINTTEAEFAAASVQRYELNMKALNLNSESLENSSVEKSSAHHHTKSHSIPKHNEEVKTETHGEEEDAHDKKPHASKDAHELKKKTEVKKEDAKQDRNEKVIQEPQATVLPVVDKKEPEESVEENTSKTSSPSPSPPAAKSWSAIASDAIKSRQASNKTVSGSMVTKTPISGTTAGVSSTNMAAATIGKSSSPLLSKQPQKKDKKYVPPSTKGIEPLGSIALRMCFDPDFISYVLRNKDVENKIPVHSIIPRGIINRANICFMSSVLQVLLYCKPFIDVINVLSTRNTNSRVGTSSCKLLDACLTMYKQFDKETYEKKFLENADDAEKTTESDAKKSSKSKSFQHCATADAVKPDEFYKTLSTIPKFKDLQWGHQEDAEEFLTHLLDQLHEELISAIDGLTDNEIQNMLQSINDEQLKVFFIRNLSRYGKAEFIKNASPRLKELIEKYGVINDDSTEENGWHEVSGSSKRGKKTKTAAKRTVEIVPSPISKLFGGQFRSVLDIPNNKESQSITLDPFQTIQLDISDAGVNDLETAFKKFSEYELLPFKSSSGNDVEAKKQTFIDKLPQVLLIQFKRFSFINNVNKDNAMTNYNAYNGRIEKIRKKIKYGHELIIPEESMSSITLKNNTSGIDDRRYKLTGVIYHHGVSSDGGHYTADVYHSEHNKWYRIDDVNITELEDDDVLKGGEEASDSRTAYILMYQKRN 337FD05527C5542D 337FD05527C5542D bre5-ubp3-11 1 6 unspecified role unspecified role neutral component neutral component purified purified 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) 5 unspecified role unspecified role neutral component neutral component purified purified 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) 9 11 direct interaction direct interaction 1.69A Rworking 0.207 Rfree 0.225 Table 1 ubp3-bre5-9 2 6 unspecified role unspecified role bait bait purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 Q01477:p.Leu208Ala 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 L A Wed May 02 16:53:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu208ala' Tue Jun 30 18:32:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu208Ala' Q01477:p.Phe209Ala 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 F A Wed May 02 20:31:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe209ala' Tue Jun 30 17:36:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe209Ala' Q01477:p.Ile210Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain I A Wed May 02 15:08:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile210ala' Tue Jun 30 17:30:18 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile210Ala' Q01477:p.Asn211Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain N A Wed May 02 15:47:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn211ala' Tue Jun 30 17:53:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn211Ala' Q01477:p.Thr212Ala 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 T A Wed May 02 17:29:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr212ala' Tue Jun 30 15:58:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr212Ala' Q01477:p.Glu216Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Wed May 02 15:49:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu216ala' Tue Jun 30 15:39:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu216Ala' Q01477:p.Phe217Ala 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 F A Wed May 02 14:58:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe217ala' Tue Jun 30 17:42:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe217Ala' Q01477:p.Glu214Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Fri Dec 08 14:13:02 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'arg214ala' Wed May 02 18:20:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu214ala' Tue Jun 30 17:02:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu214Ala' Q01477:p.Leu208_Phe209delinsAlaAla mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain LF AA Fri Dec 08 14:04:14 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'leu208ala,phe209ala' Wed May 02 18:19:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_phe208-209ala_ala' Tue Jun 30 18:17:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu208_Phe209delinsAlaAla' Q01477:p.[Ile210Ala;Phe217Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain I A certain certain sequence position certain certain certain sequence position certain F A Wed May 02 15:51:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile210ala,phe217ala' Tue Jun 30 17:25:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile210Ala;Phe217Ala]' Q01477:p.[Asn211Ala;Phe217Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain N A certain certain sequence position certain certain certain sequence position certain F A Wed May 02 18:11:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn211ala,phe217ala' Tue Jun 30 16:54:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn211Ala;Phe217Ala]' Q01477:p.Leu208_Asn211delinsAlaAlaAlaAla mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain LFIN AAAA Fri Dec 08 14:23:44 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'leu208ala,phe209ala,ile210ala,asn211ala' Wed May 02 18:23:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_phe_ile_asn208-211ala_ala_ala_ala' Tue Jun 30 18:48:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu208_Asn211delinsAlaAlaAlaAla' Q01477:p.[Leu208_Asn211delinsAlaAlaAlaAla;Phe217Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain LFIN AAAA certain certain sequence position certain certain certain sequence position certain F A Fri Dec 08 14:06:03 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'leu208ala,phe209ala,ile210ala,asn211ala,phe217ala' Wed May 02 16:46:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_phe_ile_asn208-211ala_ala_ala_ala,phe217ala' Tue Jun 30 18:18:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu208_Asn211delinsAlaAlaAlaAla;Phe217Ala]' Q01477:p.Arg224Ala 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 R A Tue Jun 30 18:09:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg224Ala' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey purified purified 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 P53741:p.Tyr17Ala 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 Y A Wed May 02 18:17:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr17ala' Tue Jun 30 19:22:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr17Ala' P53741:p.Lys80Ala 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 K A Wed May 02 19:20:51 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys80ala' Tue Jun 30 15:39:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys80Ala' P53741:p.Lys82Ala 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 K A Wed May 02 19:55:45 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys82ala' Tue Jun 30 18:31:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys82Ala' P53741:p.Glu105Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain E A Wed May 02 18:03:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu105ala' Tue Jun 30 19:36:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu105Ala' ecoli Escherichia coli (strain K12) 14 31 direct interaction direct interaction The lys80ala or lys82ala mutation in BRE5 disrupted binding in combination with a ile210ala mutation of UBP3. The tyr17ala,lys82ala mutation in BRE5 disrupted binding in combination with a ile210ala mutation of UBP3 Fig. 4A, 4B We initially prepared single alanine replacements of several Bre5 and Ubp3 residues shown to mediate protein-protein interactions in the complex and used GST-pulldown studies to compare the protein binding properties of these mutants. bre5-ubp3-12 4 6 unspecified role unspecified role neutral component neutral component purified purified c-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q01477:p.[Leu208_Asn211delinsAlaAlaAlaAla;Phe217Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain LFIN AAAA certain certain sequence position certain certain certain sequence position certain F A Fri Dec 08 14:36:07 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'leu208ala,phe209ala,ile210ala,asn211ala,phe217ala' Wed May 02 20:03:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_phe_ile_asn208-211ala_ala_ala_ala,phe217ala' Tue Jun 30 19:10:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu208_Asn211delinsAlaAlaAlaAla;Phe217Ala]' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component purified purified 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 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) 40 44 direct interaction direct interaction Fig 4C ubp3-bre5-10 3 6 unspecified role unspecified role bait bait purified purified region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey purified purified 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 P53741:p.[Tyr17Ala;Lys82Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Y A certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:37:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr17ala,lys82ala' Tue Jun 30 15:46:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr17Ala;Lys82Ala]' ecoli Escherichia coli (strain K12) physical association physical association Fig. 4D bre5-ubp3-14 3 6 unspecified role unspecified role prey prey purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q01477:p.Leu208_Asn211delinsAlaAlaAlaAla mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain LFIN AAAA Fri Dec 08 14:30:27 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'leu208ala,phe209ala,ile210ala,asn211ala' Wed May 02 16:04:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_phe_ile_asn208-211ala_ala_ala_ala' Tue Jun 30 16:06:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu208_Asn211delinsAlaAlaAlaAla' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Fig. 4D