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/ http://www.ebi.ac.uk/ greschik-2008-2 chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 gal4 vp16 complement gal4 vp16 complementation karyoplasmic interaction ion strategy KISS mammalian two hybrid gal4 vp16 complement predetermined predetermined participant predetermined Greschik H., Althage M., Flaig R., Sato Y., Chavant V., Peluso-Iltis C., Choulier L., Cronet P., Rochel N., Schüle R., Strömstedt PE., Moras D. Some results from figure 3B and 5B have not been included as chimeric constructs are used, which are of dubious biological relevance. moras@igbmc.u-strasbg.fr. The Journal of biological chemistry 2008 Only protein-protein interactions imex curation imex curation greschik-2008-3 in vitro In vitro emsa electrophoretic mobility shift assay Gel retardation assay band shift band shift Gel retardation assay emsa weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Greschik H., Althage M., Flaig R., Sato Y., Chavant V., Peluso-Iltis C., Choulier L., Cronet P., Rochel N., Schüle R., Strömstedt PE., Moras D. moras@igbmc.u-strasbg.fr. The Journal of biological chemistry 2008 Only protein-protein interactions imex curation imex curation greschik-2008-1 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Greschik H., Althage M., Flaig R., Sato Y., Chavant V., Peluso-Iltis C., Choulier L., Cronet P., Rochel N., Schüle R., Strömstedt PE., Moras D. moras@igbmc.u-strasbg.fr. The Journal of biological chemistry 2008 Only protein-protein interactions imex curation imex curation prgc1_human Peroxisome proliferator-activated receptor gamma coactivator 1-alpha Ligand effect modulator 6 PPARGC1A LEM6 PGC1 PGC1A PPARGC1 protein protein human Homo sapiens Human MAWDMCNQDSESVWSDIECAALVGEDQPLCPDLPELDLSELDVNDLDTDSFLGGLKWCSDQSEIISNQYNNEPSNIFEKIDEENEANLLAVLTETLDSLPVDEDGLPSFDALTDGDVTTDNEASPSSMPDGTPPPQEAEEPSLLKKLLLAPANTQLSYNECSGLSTQNHANHNHRIRTNPAIVKTENSWSNKAKSICQQQKPQRRPCSELLKYLTTNDDPPHTKPTENRNSSRDKCTSKKKSHTQSQSQHLQAKPTTLSLPLTPESPNDPKGSPFENKTIERTLSVELSGTAGLTPPTTPPHKANQDNPFRASPKLKSSCKTVVPPPSKKPRYSESSGTQGNNSTKKGPEQSELYAQLSKSSVLTGGHEERKTKRPSLRLFGDHDYCQSINSKTEILINISQELQDSRQLENKDVSSDWQGQICSSTDSDQCYLRETLEASKQVSPCSTRKQLQDQEIRAELNKHFGHPSQAVFDDEADKTGELRDSDFSNEQFSKLPMFINSGLAMDGLFDDSEDESDKLSYPWDGTQSYSLFNVSPSCSSFNSPCRDSVSPPKSLFSQRPQRMRSRSRSFSRHRSCSRSPYSRSRSRSPGSRSSSRSCYYYESSHYRHRTHRNSPLYVRSRSRSPYSRRPRYDSYEEYQHERLKREEYRREYEKRESERAKQRERQRQKAIEERRVIYVGKIRPDTTRTELRDRFEVFGEIEECTVNLRDDGDSYGFITYRYTCDAFAALENGYTLRRSNETDFELYFCGRKQFFKSNYADLDSNSDDFDPASTKSKYDSLDFDSLLKEAQRSLRR err1_human Steroid hormone receptor ERR1 Estrogen-related receptor alpha Estrogen receptor-like 1 Nuclear receptor subfamily 3 group B member 1 ESRRA ERR1 ESRL1 NR3B1 protein protein human Homo sapiens Human MSSQVVGIEPLYIKAEPASPDSPKGSSETETEPPVALAPGPAPTRCLPGHKEEEDGEGAGPGEQGGGKLVLSSLPKRLCLVCGDVASGYHYGVASCEACKAFFKRTIQGSIEYSCPASNECEITKRRRKACQACRFTKCLRVGMLKEGVRLDRVRGGRQKYKRRPEVDPLPFPGPFPAGPLAVAGGPRKTAAPVNALVSHLLVVEPEKLYAMPDPAGPDGHLPAVATLCDLFDREIVVTISWAKSIPGFSSLSLSDQMSVLQSVWMEVLVLGVAQRSLPLQDELAFAEDLVLDEEGARAAGLGELGAALLQLVRRLQALRLEREEYVLLKALALANSDSVHIEDAEAVEQLREALHEALLEYEAGRAGPGGGAERRRAGRLLLTLPLLRQTAGKVLAHFYGVKLEGKVPMHKLFLEMLEAMMD esr1_human Estrogen receptor ESR1 ESR NR3A1 Estradiol receptor ER-alpha Nuclear receptor subfamily 3 group A member 1 protein protein human Homo sapiens Human MTMTLHTKASGMALLHQIQGNELEPLNRPQLKIPLERPLGEVYLDSSKPAVYNYPEGAAYEFNAAAAANAQVYGQTGLPYGPGSEAAAFGSNGLGGFPPLNSVSPSPLMLLHPPPQLSPFLQPHGQQVPYYLENEPSGYTVREAGPPAFYRPNSDNRRQGGRERLASTNDKGSMAMESAKETRYCAVCNDYASGYHYGVWSCEGCKAFFKRSIQGHNDYMCPATNQCTIDKNRRKSCQACRLRKCYEVGMMKGGIRKDRRGGRMLKHKRQRDDGEGRGEVGSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV dr0_probe direct repeat oligo with 0 bp spacing ds dna double stranded deoxyribonucleic acid ds DNA ds dna chemical synthesis Chemical synthesis (Chemical synthesis) AGCTTCAGGTCAAGGTCAGAGAGCT ere_probe oestrogen response element ds dna double stranded deoxyribonucleic acid ds DNA ds dna chemical synthesis Chemical synthesis (Chemical synthesis) GATCCGTCAGGTCACAGTGACCTGATACATC esrra-ppargc1a-1 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.Leu144Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.Leu210Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.[Leu144Ala;Leu210Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false box3 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false box2 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 2, 4 ppargc1a-esrra-1 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Q9UBK2:p.Leu144Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false box2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 2-2 ppargc1a-esrra-2 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Q9UBK2:p.Leu211Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.Tyr213Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.Cys207Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.Cys207Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.Leu210Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false box3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 2-3, figure 3A These results revealed that amino acids at positions -6 to -3 (Arg-204/Arg-205 and Cys-207) determine high affinity binding of the PGC-1a box3 region to the ERRa LBD. ppargc1a-esrra-3 3358427 N/A 3358431 unspecified role unspecified role prey prey region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Q9UBK2:p.Pro202_Gln203delinsAlaAla 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 false Q9UBK2:p.Arg204_Arg205delinsAlaAla mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false truncated_box3 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false figure 3A-2 These results revealed that amino acids at positions -6 to -3 (Arg-204/Arg-205 and Cys-207) determine high affinity binding of the PGC-1a box3 region to the ERRa LBD. ppargc1a-esrra-4 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false box3 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey P11474:p.[Met258Val;Gln262Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P11474:p.His341Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp423Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Met421_Asp423del mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 4A ppargc1a-esrra-6 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed box2 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey P11474:p.[Met258Val;Gln262Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P11474:p.His341Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp423Ala 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 false P11474:p.Met421_Asp423del mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 4Ai ppargc1a-esrra-5 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed box2 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false box3 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey P11474:p.Asp338_His341delinsGlyValTyrThrPheLeuSerSerThrLeuLysSer mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Arg315Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P11474:p.Ser259His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 4C1 ppargc1a-esrra-7 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed Q9UBK2:p.Lys212Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false box3 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.Leu211Ala mutation mutation certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 3B ppargc1a-esrra-8 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey P11474:p.Glu343Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 4C-2 ppargc1a-esrra-9 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed box2 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false box3 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 3358431 unspecified role unspecified role prey prey P11474:p.Glu343Ala 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 false region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 4C-3 ppargc1a-esrra-10 3358427 N/A 3358430 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false box2 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false N/A 3358432 unspecified role unspecified role prey prey region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P03372:p.[Val376Met;Glu380Gln] mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Figure 5D The wild-type ERa LBD interacted in an estradiol (E2)-dependent manner, detectable only with the PGC-1a ID and box2 but not with box3. E2- estradiol CHEBI:23965 qwertyuiop-1 3358428 N/A 3358433 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified P11474:p.Arg315Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Glu343Ala 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 false P11474:p.Asp338_His341delinsGlyValTyrThrPheLeuSerSerThrLeuLysSer mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 2.0 direct interaction direct interaction false false false Figure 6 A/B, supp fig 2B dr0_probe-esrra-1 3358428 N/A 3358433 competitor competitor competitor unspecified role unspecified role purified purified chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358433 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified in vitro In vitro Stoichiometry: 2.0 physical association physical association false false false Figure 6A-2 dr0_probe-esrra-2 3358428 N/A 3358433 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified P11474:p.Arg315Ala mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338Ala mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain false P11474:p.Glu343Ala mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338_His341delinsGlyValTyrThrPheLeuSerSerThrLeuLysSer mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 1.0 direct interaction direct interaction false false false Figure 6 A/B, supp fig 2B Only a weak band is seen here for the DR0-E343A ERRa mutant (monomer). dr0_probe-esrra-3 3358428 N/A 3358434 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified P11474:p.Arg315Ala 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 false P11474:p.Asp338Ala 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 false P11474:p.Glu343Ala 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 false P11474:p.Asp338_His341delinsGlyValTyrThrPheLeuSerSerThrLeuLysSer mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 2.0 direct interaction direct interaction false false false Figure 6 A/C ere_probe-esrra-1 3358428 N/A 3358434 competitor competitor competitor unspecified role unspecified role purified purified chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358434 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified in vitro In vitro Stoichiometry: 2.0 direct interaction direct interaction false false false Figure 6A-ii ere_probe-esrra-2 3358428 N/A 3358430 unspecified role unspecified role prey prey partially purified partially purified ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.[Leu144Ala;Leu210Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) N/A 3358433 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified P11474:p.Arg315Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Glu343Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338_His341delinsGlyValTyrThrPheLeuSerSerThrLeuLysSer mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 2.0 physical association physical association false false false Figure 6B-1, supp fig 2B The addition of small amounts of E. coli-expressed, partially purified GST-PGC-1a ID fusion protein resulted in an upshift of DNA-bound ERRa (Fig. 6, B and C). This upshift was not observed when GST-PGC-1a ID(L144A/L210A) was added. Compared with the wild-type receptor, all DNA-bound mutants were less efficiently shifted by GST-PGC-1a ID. Mutations in the H8–H9 loop such as R315A or D338A influence not only homodimerization but also Coactivator binding of DNA-bound, full-length ERRa. dr0_probe-esrra-4 3358428 N/A 3358430 inhibitor inhibitor prey prey partially purified partially purified ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3358433 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified P11474:p.Arg315Ala mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338Ala mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain false P11474:p.Glu343Ala mutation causing mutation causing an interaction mutation enabling interaction mutation causing mutation causing an interaction certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 1.0 direct interaction direct interaction false false false Figure 6B, supp fig 2B Only a very weak band is seen here for the DR0-E343A ERRa mutant (monomer). DNA-bound ERRa(SwapH8–H9) did not interact with GST-PGC-1a ID. ere_probe-esrra-3 3358428 N/A 3358430 unspecified role unspecified role prey prey partially purified partially purified ID_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UBK2:p.[Leu144Ala;Leu210Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) N/A 3358434 unspecified role unspecified role bait bait purified purified region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 3358431 unspecified role unspecified role prey prey partially purified partially purified P11474:p.Arg315Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Glu343Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P11474:p.Asp338_His341delinsGlyValTyrThrPheLeuSerSerThrLeuLysSer mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 2.0 physical association physical association false false false Figure 6C The addition of small amounts of E. coli-expressed, partially purified GST-PGC-1a ID fusion protein resulted in an upshift of DNA-bound ERRa (Fig. 6, B and C). This upshift was not observed when GST-PGC-1a ID(L144A/L210A) was added. Compared with the wild-type receptor, all DNA-bound mutants were less efficiently shifted by GST-PGC-1a ID. Mutations in the H8–H9 loop such as R315A or D338A influence not only homodimerization but also Coactivator binding of DNA-bound, full-length ERRa. 12345rtyuuuu-1 3358429 N/A 3358430 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 false Stoichiometry: 1.0 N/A 3358431 unspecified role unspecified role neutral component neutral component purified purified n-terminus 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 false region_including_nr_lbd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Table 1 2.11 A RWork = 0.212 RFree = 0.255