UniProt UniProt http://www.uniprot.org Structural basis for membrane insertion by the human ER membrane protein complex. Structural basis for membrane insertion by the human ER membrane protein complex. Science (New York, N.Y.) 2020 imex curation Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. voorhees@caltech.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. EMC8 and 9 are functional paralogs, and their binding to EMC2 is mutually exclusive. Each indivdiual ECM complex will, therefore, only contain one of these molecules. The human EMC was affinity purified using an immobilized, protease-cleavable, GFP-nanobody and reconstituted into lipid nanodiscs prior to cryo-electron microscopy. voorhees@caltech.edu Accepted 2020-AUG-27 AT 13:11 BST AT 13:11 BST by KALPANAP Science (New York, N.Y.) 2020 Only protein-protein interactions imex curation imex curation Structural basis for membrane insertion by the human ER membrane protein complex. Structural basis for membrane insertion by the human ER membrane protein complex. Science (New York, N.Y.) 2020 imex curation Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. voorhees@caltech.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. voorhees@caltech.edu Accepted 2020-AUG-27 AT 13:11 BST AT 13:11 BST by KALPANAP Science (New York, N.Y.) 2020 Only protein-protein interactions imex curation imex curation Structural basis for membrane insertion by the human ER membrane protein complex. Structural basis for membrane insertion by the human ER membrane protein complex. Science (New York, N.Y.) 2020 imex curation Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. voorhees@caltech.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation rabit-reticulocyte Oryctolagus cuniculus reticulocyte reticulocyte Reticulocytes anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. voorhees@caltech.edu Accepted 2020-AUG-27 AT 13:11 BST AT 13:11 BST by KALPANAP Science (New York, N.Y.) 2020 Only protein-protein interactions imex curation imex curation Structural basis for membrane insertion by the human ER membrane protein complex. Structural basis for membrane insertion by the human ER membrane protein complex. Science (New York, N.Y.) 2020 imex curation Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. voorhees@caltech.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro tem transmission electron microscopy tem structure determination identification by structure determination identification by structure determination structure determination Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. EMC8 and 9 are functional paralogs, and their binding to EMC2 is mutually exclusive. Each indivdiual ECM complex will, therefore, only contain one of these molecules. The human EMC was affinity purified using an immobilized, protease-cleavable, GFP-nanobody and reconstituted into lipid nanodiscs prior to cryo-electron microscopy. voorhees@caltech.edu Accepted 2020-AUG-27 AT 13:11 BST AT 13:11 BST by KALPANAP Science (New York, N.Y.) 2020 Only protein-protein interactions imex curation imex curation Structural basis for membrane insertion by the human ER membrane protein complex. Structural basis for membrane insertion by the human ER membrane protein complex. Science (New York, N.Y.) 2020 imex curation Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. voorhees@caltech.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip ms participant Identification by mass spectrometry ms participant Pleiner T., Pinton Tomaleri G., Januszyk K., Inglis AJ., Hazu M., Voorhees RM. EMC8 and 9 are functional paralogs, and their binding to EMC2 is mutually exclusive. Each indivdiual ECM complex will, therefore, only contain one of these molecules. voorhees@caltech.edu Accepted 2020-AUG-27 AT 13:11 BST AT 13:11 BST by KALPANAP Science (New York, N.Y.) 2020 Only protein-protein interactions imex curation imex curation emc5_human ER membrane protein complex subunit 5 MMGT1 TMEM32 Transmembrane protein 32 EMC5 Membrane magnesium transporter 1 protein protein human Homo sapiens Human MAPSLWKGLVGIGLFALAHAAFSAAQHRSYMRLTEKEDESLPIDIVLQTLLAFAVTCYGIVHIAGEFKDMDATSELKNKTFDTLRNHPSFYVFNHRGRVLFRPSDTANSSNQDALSSNTSLKLRKLESLRR DF6255A8172C6016 emc3_human ER membrane protein complex subunit 3 EMC3 TMEM111 Transmembrane protein 111 protein protein human Homo sapiens Human MAGPELLLDSNIRLWVVLPIVIITFFVGMIRHYVSILLQSDKKLTQEQVSDSQVLIRSRVLRENGKYIPKQSFLTRKYYFNNPEDGFFKKTKRKVVPPSPMTDPTMLTDMMKGNVTNVLPMILIGGWINMTFSGFVTTKVPFPLTLRFKPMLQQGIELLTLDASWVSSASWYFLNVFGLRSIYSLILGQDNAADQSRMMQEQMTGAAMAMPADTNKAFKTEWEALELTDHQWALDDVEEELMAKDLHFEGMFKKELQTSIF 0FB279E54D6C1FCC emc10_human ER membrane protein complex subunit 10 EMC10 C19orf63 Hematopoietic signal peptide-containing membrane domain-containing protein 1 INM02 UNQ764/PRO1556 protein protein human Homo sapiens Human MAAASAGATRLLLLLLMAVAAPSRARGSGCRAGTGARGAGAEGREGEACGTVGLLLEHSFEIDDSANFRKRGSLLWNQQDGTLSLSQRQLSEEERGRLRDVAALNGLYRVRIPRRPGALDGLEAGGYVSSFVPACSLVESHLSDQLTLHVDVAGNVVGVSVVTHPGGCRGHEVEDVDLELFNTSVQLQPPTTAPGPETAAFIERLEMEQAQKAKNPQEQKSFFAKYWMYIIPVVLFLMMSGAPDTGGQGGGGGGGGGGGSGR 816940C105B95AA1 emc7_human ER membrane protein complex subunit 7 EMC7 C11orf3 C15orf24 HT022 UNQ905/PRO1926 protein protein human Homo sapiens Human MAAALWGFFPVLLLLLLSGDVQSSEVPGAAAEGSGGSGVGIGDRFKIEGRAVVPGVKPQDWISAARVLVDGEEHVGFLKTDGSFVVHDIPSGSYVVEVVSPAYRFDPVRVDITSKGKMRARYVNYIKTSEVVRLPYPLQMKSSGPPSYFIKRESWGWTDFLMNPMVMMMVLPLLIFVLLPKVVNTSDPDMRREMEQSMNMLNSNHELPDVSEFMTRLFSSKSSGKSSSGSSKTGKSGAGKRR A71930B89A4C2458 emc2_human ER membrane protein complex subunit 2 EMC2 KIAA0103 TTC35 Tetratricopeptide repeat protein 35 protein protein human Homo sapiens Human MAKVSELYDVTWEEMRDKMRKWREENSRNSEQIVEVGEELINEYASKLGDDIWIIYEQVMIAALDYGRDDLALFCLQELRRQFPGSHRVKRLTGMRFEAMERYDDAIQLYDRILQEDPTNTAARKRKIAIRKAQGKNVEAIRELNEYLEQFVGDQEAWHELAELYINEHDYAKAAFCLEELMMTNPHNHLYCQQYAEVKYTQGGLENLELSRKYFAQALKLNNRNMRALFGLYMSASHIASNPKASAKTKKDNMKYASWAASQINRAYQFAGRSKKETKYSLKAVEDMLETLQITQS CA5903AEC3B3E36A emc6_human ER membrane protein complex subunit 6 EMC6 TMEM93 Transmembrane protein 93 protein protein human Homo sapiens Human MAAVVAKREGPPFISEAAVRGNAAVLDYCRTSVSALSGATAGILGLTGLYGFIFYLLASVLLSLLLILKAGRRWNKYFKSRRPLFTGGLIGGLFTYVLFWTFLYGMVHVY 5120B6E4CC12FD48 emc1_human ER membrane protein complex subunit 1 EMC1 KIAA0090 PSEC0263 protein protein human Homo sapiens Human MAAEWASRFWLWATLLIPAAAVYEDQVGKFDWRQQYVGKVKFASLEFSPGSKKLVVATEKNVIAALNSRTGEILWRHVDKGTAEGAVDAMLLHGQDVITVSNGGRIMRSWETNIGGLNWEITLDSGSFQALGLVGLQESVRYIAVLKKTTLALHHLSSGHLKWVEHLPESDSIHYQMVYSYGSGVVWALGVVPFSHVNIVKFNVEDGEIVQQVRVSTPWLQHLSGACGVVDEAVLVCPDPSSRSLQTLALETEWELRQIPLQSLDLEFGSGFQPRVLPTQPNPVDASRAQFFLHLSPSHYALLQYHYGTLSLLKNFPQTALVSFATTGEKTVAAVMACRNEVQKSSSSEDGSMGSFSEKSSSKDSLACFNQTYTINLYLVETGRRLLDTTITFSLEQSGTRPERLYIQVFLKKDDSVGYRALVQTEDHLLLFLQQLAGKVVLWSREESLAEVVCLEMVDLPLTGAQAELEGEFGKKADGLLGMFLKRLSSQLILLQAWTSHLWKMFYDARKPRSQIKNEINIDTLARDEFNLQKMMVMVTASGKLFGIESSSGTILWKQYLPNVKPDSSFKLMVQRTTAHFPHPPQCTLLVKDKESGMSSLYVFNPIFGKWSQVAPPVLKRPILQSLLLPVMDQDYAKVLLLIDDEYKVTAFPATRNVLRQLHELAPSIFFYLVDAEQGRLCGYRLRKDLTTELSWELTIPPEVQRIVKVKGKRSSEHVHSQGRVMGDRSVLYKSLNPNLLAVVTESTDAHHERTFIGIFLIDGVTGRIIHSSVQKKAKGPVHIVHSENWVVYQYWNTKARRNEFTVLELYEGTEQYNATAFSSLDRPQLPQVLQQSYIFPSSISAMEATITERGITSRHLLIGLPSGAILSLPKALLDPRRPEIPTEQSREENLIPYSPDVQIHAERFINYNQTVSRMRGIYTAPSGLESTCLVVAYGLDIYQTRVYPSKQFDVLKDDYDYVLISSVLFGLVFATMITKRLAQVKLLNRAWR C61B67802052A8A7 emc4_human ER membrane protein complex subunit 4 EMC4 Cell proliferation-inducing gene 17 protein TMEM85 Transmembrane protein 85 HSPC184 PIG17 protein protein human Homo sapiens Human MTAQGGLVANRGRRFKWAIELSGPGGGSRGRSDRGSGQGDSLYPVGYLDKQVPDTSVQETDRILVEKRCWDIALGPLKQIPMNLFIMYMAGNTISIFPTMMVCMMAWRPIQALMAISATFKMLESSSQKFLQGLVYLIGNLMGLALAVYKCQSMGLLPTHASDWLAFIEPPERMEFSGGGLLL 96888D46FF22DEDE emc9_human ER membrane protein complex subunit 9 EMC9 C14orf122 FAM158A Protein FAM158A CGI-112 protein protein human Homo sapiens Human MGEVEISALAYVKMCLHAARYPHAAVNGLFLAPAPRSGECLCLTDCVPLFHSHLALSVMLEVALNQVDVWGAQAGLVVAGYYHANAAVNDQSPGPLALKIAGRIAEFFPDAVLIMLDNQKLVPQPRVPPVIVLENQGLRWVPKDKNLVMWRDWEESRQMVGALLEDRAHQHLVDFDCHLDDIRQDWTNQRLNTQITQWVGPTNGNGNA 173647F7DFF63510 emc8_human ER membrane protein complex subunit 8 EMC8 C16orf2 C16orf4 COX4AL COX4NB FAM158B NOC4 Neighbor of COX4 Protein FAM158B protein protein human Homo sapiens Human MPGVKLTTQAYCKMVLHGAKYPHCAVNGLLVAEKQKPRKEHLPLGGPGAHHTLFVDCIPLFHGTLALAPMLEVALTLIDSWCKDHSYVIAGYYQANERVKDASPNQVAEKVASRIAEGFSDTALIMVDNTKFTMDCVAPTIHVYEHHENRWRCRDPHHDYCEDWPEAQRISASLLDSRSYETLVDFDNHLDDIRNDWTNPEINKAVLHLC 41367FDD98769250 emc1-emc10-1 4 6 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 7 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 9 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 15 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 13 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 8 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 12 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 11 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 10 unspecified role unspecified role neutral component neutral component 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 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. physical association physical association Fig. 1, figs. S2 and S3, and table S1 emc1-emc10-2 5 6 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 14 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 15 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 13 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 association association Supp Fig 1c (top panel) emc2-emc1-2 2 15 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed 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 association association Supp Fig 1c (bottom panel) emc1-emc10-3 1 6 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 14 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 7 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 9 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 15 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 13 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 8 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 10 unspecified role unspecified role neutral component neutral component 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 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 12 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 11 unspecified role unspecified role neutral component neutral component purified purified human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. physical association physical association Supp Fig. 1D emc6-mmgt1-4 3 6 unspecified role unspecified role bait bait in vitro translated in vitro translated protein in vitro translated region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey in vitro translated in vitro translated protein in vitro translated Q9BV81:p.[Gly45Tyr;Gly48Ala] 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 G Y certain certain sequence position certain certain certain sequence position certain G A Tue Jun 30 17:46:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gly45Tyr;Gly48Ala]' region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9BV81:p.[Val33Lys;Leu36Lys] 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 V K certain certain sequence position certain certain certain sequence position certain L K Tue Jun 30 18:45:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Val33Lys;Leu36Lys]' physical association physical association Fig. S6c emc2-emc8-5 3 10 unspecified role unspecified role unspecified role unspecified role cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q15006:p.Tyr200Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y A Q15006:p.Arg227Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Q15006:p.Arg28Ala 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 Q15006:p.Glu156Ala 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 Q15006:p.Glu160Ala 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 Q15006:p.Glu180Ala 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 Q15006:p.Trp259Ala 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 W A Q15006:p.Tyr171Ala 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 15 unspecified role unspecified role bait bait cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 2C emc3-emc2-1 3 10 unspecified role unspecified role unspecified role unspecified role cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q15006:p.Tyr200Ala 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 Q15006:p.Arg227Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Q15006:p.Arg28Ala 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 Q15006:p.Glu156Ala 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 Q15006:p.Glu160Ala 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 Q15006:p.Glu180Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain E A Q15006:p.Trp259Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain W A Q15006:p.Tyr171Ala 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 7 unspecified role unspecified role bait bait cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 2C mmgt1-emc2-1 3 6 unspecified role unspecified role bait bait cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role unspecified role unspecified role cell lysate cell lysate in vitro translated in vitro translated protein in vitro translated region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q15006:p.Tyr200Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y A Q15006:p.Arg227Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Q15006:p.Arg28Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain R A Q15006:p.Glu156Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain E A Q15006:p.Glu160Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain E A Q15006:p.Glu180Ala 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 Q15006:p.Trp259Ala 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 W A Q15006:p.Tyr171Ala 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 physical association physical association Fig. 2C