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 structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. Cell (0092-8674) 2006 imex curation 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 01-DEC-2006: Contacted by JYOTI. orchard Only protein-protein interactions 5 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 Accepted 2006-NOV-21 by ORCHARD. 01-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 orchard Only protein-protein interactions 5 imex curation The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. Cell (0092-8674) 2006 imex curation 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 01-DEC-2006: Contacted by JYOTI. orchard Only protein-protein interactions 5 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 Accepted 2006-NOV-20 by ORCHARD. 01-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 orchard Only protein-protein interactions 5 imex curation The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. Cell (0092-8674) 2006 imex curation 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 01-DEC-2006: Contacted by JYOTI. orchard Only protein-protein interactions 5 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 Accepted 2006-NOV-21 by ORCHARD. 01-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 orchard Only protein-protein interactions 5 imex curation The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. The structure of the zetazeta transmembrane dimer reveals features essential for its assembly with the T cell receptor. Cell (0092-8674) 2006 imex curation 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 01-DEC-2006: Contacted by JYOTI. orchard Only protein-protein interactions 5 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 Accepted 2006-NOV-21 by ORCHARD. 01-DEC-2006: Contacted by JYOTI. Cell (0092-8674) 2006 orchard Only protein-protein interactions 5 imex curation 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 9408260374856EE9 cd247-cd247 2 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain cys32 forms an interchain disulfide bond biological feature biological feature certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 2h label 2h label deuterium D2 2H2 2H2 deuterium D2 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined cys32 forms an interchain disulfide bond biological feature biological feature certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 14 9 direct interaction direct interaction 2, Table 1 The portion of the zzTM making interchain contacts (colored yellow and green) comprises almost the entire TM domain (20 of 23 TM residues cd247-cd247-1 1 5 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 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 D A Wed May 02 18:54:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp36ala' Tue Jun 30 19:30:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp36Ala' 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 L A Wed May 02 18:06:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu39ala' Tue Jun 30 20:17:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu39Ala' 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 Y A Wed May 02 15:10:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr42ala' Tue Jun 30 17:07:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr42Ala' 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 T A Wed May 02 18:52:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr47ala' Tue Jun 30 15:41:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr47Ala' 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 F A Wed May 02 15:54:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe50ala' Tue Jun 30 20:05:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe50Ala' in vitro In vitro physical association physical association 3 B substitutions at positions D6, L9, Y12, T17, and F20 (Figures 3B and 3C) resulted in impaired dimer formation cd247-cd247-2 3 5 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 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 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 Y A Wed May 02 19:30:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr42ala' Tue Jun 30 18:04:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr42Ala' 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 Y F Wed May 02 15:30:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr42phe' Tue Jun 30 16:07:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr42Phe' 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 T A Wed May 02 14:28:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr47ala' Tue Jun 30 18:32:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr47Ala' P20963:p.Asp36Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D N Wed May 02 17:56:29 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp36asn' Tue Jun 30 20:04:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp36Asn' P20963:p.Asp36Ser mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D S Wed May 02 15:31:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp36ser' Tue Jun 30 18:38:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp36Ser' P20963:p.Asp36Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D A Wed May 02 19:55:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp36ala' 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.Asp36Ala' in vitro In vitro physical association physical association 3 C, 4C substitutions at positions D6, L9, Y12, T17, and F20 (Figures 3B and 3C) resulted in impaired dimer formation Substitution of either Y12 or T17 results in a dramatic reduction of covalent zz dimer formation. 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. the initial rate of T17A dimer formation was found to be 25-fold lower than WT even though the disulfide- bonded dimer accumulated to 30% of WT by 60 min cd247-cd247-3 4 5 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 D A Wed May 02 16:45:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp36ala' Tue Jun 30 17:48:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp36Ala' 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 Y A Wed May 02 15:27:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr42ala' Tue Jun 30 20:01:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr42Ala' 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 Y F Wed May 02 16:21:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr42phe' Tue Jun 30 19:48:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr42Phe' 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 T A Wed May 02 18:05:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr47ala' Tue Jun 30 15:57:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr47Ala' 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 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 in vitro In vitro 5 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 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 in vitro In vitro physical association physical association 3 D, E, 4 D Efficient dimerization was observed for mixed dimers of one WT and one mutant chain with a different polar residue (E, N, or S) at position 6 (Figure 3D, lanes 2�4), and only the mixed dimer between a WT and a D6A mutant chain showed impaired dimer formation A kinetic analysis of this D-A combination (Figure 3E) indicated that while the D-A mixed dimer accumulated to 60% of WT by 60 min, the initial rate of dimer formation was reduced 8- to 10-fold compared to WT. 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. Substitution of either Y12 or T17 results in a dramatic reduction of covalent zz dimer formation. cd247-cd247-4 4 5 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 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 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 Y T Wed May 02 20:28:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr42thr' Tue Jun 30 17:03:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr42Thr' in vitro In vitro 5 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 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 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 T Y Wed May 02 17:32:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr47tyr' Tue Jun 30 17:55:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr47Tyr' in vitro In vitro physical association physical association 4 D