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/ call-2006-4 The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Call ME., Schnell JR., Xu C., Lutz RA., Chou JJ., Wucherpfennig KW. Two-step sequential nondenaturing IP targeting PC-tagged zeta mutant strand followed by the HA-tagged zeta wild-type strand was carried out. After primary IP, nondenaturing elution with EDTA (PC tag) was carried out and eluted complexes were incubated with subsequent antibodies. james_chou@hms.harvard.edu,kai_wucherpfennig@dfci.harvard.edu Cell (0092-8674) 2006 Only protein-protein interactions 5 imex curation Dimers consisting of two zeta transmembrane region peptides were reconstituted in detergent micelles for solution NMR. To distinguish between interchain and intrachain NOEs one peptide was 15N-labeled and fully deuterated while the other peptide was unlabeled. call-2006-1 The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Call ME., Schnell JR., Xu C., Lutz RA., Chou JJ., Wucherpfennig KW. Dimers consisting of two zeta transmembrane region peptides were reconstituted in detergent micelles for solution NMR. To distinguish between interchain and intrachain NOEs one peptide was 15N-labeled and fully deuterated while the other peptide was unlabeled. james_chou@hms.harvard.edu,kai_wucherpfennig@dfci.harvard.edu Cell (0092-8674) 2006 Only protein-protein interactions 5 imex curation call-2006-2 The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel predetermined predetermined participant predetermined Call ME., Schnell JR., Xu C., Lutz RA., Chou JJ., Wucherpfennig KW. The mRNAs encoding the wild-type or mutant zeta chain were cotranslationally translocated into purified ER microsomes and allowed to fold and assemble under conditions similar to those found in cells. The ER-associated proteins were separated by nonreducing SDS-PAGE. james_chou@hms.harvard.edu,kai_wucherpfennig@dfci.harvard.edu Cell (0092-8674) 2006 Only protein-protein interactions 5 imex curation Dimers consisting of two zeta transmembrane region peptides were reconstituted in detergent micelles for solution NMR. To distinguish between interchain and intrachain NOEs one peptide was 15N-labeled and fully deuterated while the other peptide was unlabeled. call-2006-3 The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. in vitro In vitro comigration in gel comigration in gel electrophoresis comigration in gel predetermined predetermined participant predetermined Call ME., Schnell JR., Xu C., Lutz RA., Chou JJ., Wucherpfennig KW. The mRNAs encoding the wild-type or mutant zeta chain were cotranslationally translocated into purified ER microsomes and allowed to fold and assemble under conditions similar to those found in cells. Proteins were isolated from digitonin lysates by IP with an antibody to a C-terminal protein C tag. james_chou@hms.harvard.edu,kai_wucherpfennig@dfci.harvard.edu Cell (0092-8674) 2006 Only protein-protein interactions 5 imex curation Dimers consisting of two zeta transmembrane region peptides were reconstituted in detergent micelles for solution NMR. To distinguish between interchain and intrachain NOEs one peptide was 15N-labeled and fully deuterated while the other peptide was unlabeled. cd3z_human T-cell surface glycoprotein CD3 zeta chain T-cell receptor T3 zeta chain CD247 CD3Z T3Z TCRZ protein protein human Homo sapiens Human MKWKALFTAAILQAQLPITEAQSFGLLDPKLCYLLDGILFIYGVILTALFLRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPQRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR cd247-cd247-3 Mutational analysis of residues involved in zeta-zeta dimer formation 568461 N/A 568465 unspecified role unspecified role bait bait P20963:p.Asp36Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P20963:p.Tyr42Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Tyr42Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Thr47Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false region 35S-methionine/cysteine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal Protein C tag fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false in vitro In vitro N/A 568465 unspecified role unspecified role prey prey c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region 35S-methionine/cysteine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 3 D, E, 4 D The initial rate of the wild-type asp6ala mixed dimer formation was 8- to 10-fold less than for the wild-type, althought the mixed dimer accumulated to 60% of the wild-type by 60 min. cd247-cd247-4 Mutational analysis of residues involved in zeta-zeta dimer formation 568461 N/A 568465 unspecified role unspecified role bait bait c-terminal Protein C tag fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region 35S-methionine/cysteine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P20963:p.Tyr42Thr mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false in vitro In vitro N/A 568465 unspecified role unspecified role prey prey c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region 35S-methionine/cysteine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P20963:p.Thr47Tyr mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false in vitro In vitro physical association physical association false false false 4 D cd247-cd247 NMR solution structure of zeta-zeta (transmembrane domain) 568462 N/A 568465 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 false cys32 forms an interchain disulfide bond biological feature biological feature certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 568465 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 false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region 2h label 2h label deuterium D2 2H2 2H2 deuterium D2 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false cys32 forms an interchain disulfide bond biological feature biological feature certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 568494 568489 direct interaction direct interaction false false false 2, Table 1 cd247-cd247-1 Mutational analysis of residues involved in zeta-zeta dimer formation 568463 N/A 568465 unspecified role unspecified role neutral component neutral component region 35S-methionine/cysteine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P20963:p.Asp36Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Leu39Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Tyr42Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Thr47Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Phe50Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning 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 3 B cd247-cd247-2 Mutational analysis of residues involved in zeta-zeta dimer formation 568464 N/A 568465 unspecified role unspecified role neutral component neutral component region 35S-methionine/cysteine labeled 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminal Protein C tag fusion protein fusion protein c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P20963:p.Tyr42Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Tyr42Phe mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Thr47Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false P20963:p.Asp36Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P20963:p.Asp36Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P20963:p.Asp36Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing 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 3 C, 4C The initial rate of tyr17ala dimer formation was 25-fold lower than for the wild-type dimer, although the mutant dimer accumulated to 30% of the wild-type by 60 min.