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/ luo-2008-1 ecoli Escherichia coli (strain K12) toxcat tox-r dimerization assay toxcat predetermined predetermined participant predetermined Luo SZ., Li R., Luo SZ., Li R. Source of GYPA inferred from mutant mapping. renhao.li@uth.tmc.edu ude.cmt.htu@il.oahner Journal of molecular biology 2008 Only protein-protein interactions imex curation imex curation luo-2008-3 in vitro In vitro reverse phase chrom reverse phase chromatography reverse phase chrom ms participant Identification by mass spectrometry ms participant Luo SZ., Li R., Luo SZ., Li R. Purified TM peptides were mixed with detergents, dried and rehydrated in reaction buffer. GP1A-TM and GP1b-TM were mixed together with n-dodecyl beta-D-maltoside at a molar ratio of 1:2:200 which was taken as an indication of complex stoichiometry. The reaction products were analyzed by reverse-phase HPLC and the assignment of the each peak was assisted by mass determination. The molar ratio of GP9 was not given. renhao.li@uth.tmc.edu ude.cmt.htu@il.oahner Journal of molecular biology 2008 Only protein-protein interactions imex curation imex curation luo-2008-4 in vitro In vitro fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Luo SZ., Li R., Luo SZ., Li R. Purified TM peptides were mixed with detergents, dried and rehydrated in reaction buffer. The reaction products were analyzed by reverse-phase HPLC and the assignment of the each peak was assisted by mass determination. renhao.li@uth.tmc.edu ude.cmt.htu@il.oahner Journal of molecular biology 2008 Only protein-protein interactions imex curation imex curation luo-2008-2 in vitro In vitro comigration in sds comigration in sds page comigration in sds weight by staining molecular weight estimation by staining weight by staining Luo SZ., Li R., Luo SZ., Li R. renhao.li@uth.tmc.edu ude.cmt.htu@il.oahner Journal of molecular biology 2008 Only protein-protein interactions imex curation imex curation gp1bb_human Platelet glycoprotein Ib beta chain GP1BB Antigen CD42b-beta protein protein human Homo sapiens Human MGSGPRGALSLLLLLLAPPSRPAAGCPAPCSCAGTLVDCGRRGLTWASLPTAFPVDTTELVLTGNNLTALPPGLLDALPALRTAHLGANPWRCDCRLVPLRAWLAGRPERAPYRDLRCVAPPALRGRLLPYLAEDELRAACAPGPLCWGALAAQLALLGLGLLHALLLVLLLCRLRRLRARARARAAARLSLTDPLVAERAGTDES p02724-pro_0000012134 Glycophorin-A GYPA GPA MN sialoglycoprotein PAS-2 Sialoglycoprotein alpha protein protein human Homo sapiens Human SSTTGVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSEISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGVIGTILLISYGIRRLIKKSPSDVKPLPSPDTDVPLSSVEIENPETSDQ 20 150 gp1ba_human Platelet glycoprotein Ib alpha chain GP1BA Antigen CD42b-alpha protein protein human Homo sapiens Human MPLLLLLLLLPSPLHPHPICEVSKVASHLEVNCDKRNLTALPPDLPKDTTILHLSENLLYTFSLATLMPYTRLTQLNLDRCELTKLQVDGTLPVLGTLDLSHNQLQSLPLLGQTLPALTVLDVSFNRLTSLPLGALRGLGELQELYLKGNELKTLPPGLLTPTPKLEKLSLANNNLTELPAGLLNGLENLDTLLLQENSLYTIPKGFFGSHLLPFAFLHGNPWLCNCEILYFRRWLQDNAENVYVWKQGVDVKAMTSNVASVQCDNSDKFPVYKYPGKGCPTLGDEGDTDLYDYYPEEDTEGDKVRATRTVVKFPTKAHTTPWGLFYSWSTASLDSQMPSSLHPTQESTKEQTTFPPRWTPNFTLHMESITFSKTPKSTTEPTPSPTTSEPVPEPAPNMTTLEPTPSPTTPEPTSEPAPSPTTPEPTSEPAPSPTTPEPTSEPAPSPTTPEPTPIPTIATSPTILVSATSLITPKSTFLTTTKPVSLLESTKKTIPELDQPPKLRGVLQGHLESSRNDPFLHPDFCCLLPLGFYVLGLFWLLFASVVLILLLSWVGHVKPQALDSGQGAALTTATQTTHLELQRGRQVTVPRAWLLFLRGSLPTFRSSLFLWVRPNGRVGPLVAGRRPSALSQGRGQDLLSTVSIRYSGHSL gpix_human Platelet glycoprotein IX GP9 Glycoprotein 9 protein protein human Homo sapiens Human MPAWGALFLLWATAEATKDCPSPCTCRALETMGLWVDCRGHGLTALPALPARTRHLLLANNSLQSVPPGAFDHLPQLQTLDVTQNPWHCDCSLTYLRLWLEDRTPEALLQVRCASPSLAAHGPLGRLTGYQLGSCGWQLQASWVRPGVLWDVALVAVAALGLALLAGLLCATTEALD gp1bb-1 1852061 N/A 1852065 unspecified role unspecified role neutral component neutral component transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n-terminal toxr tag tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Stoichiometry: 2.0 physical association physical association false false false Fig. 2B gypa-1 1852061 N/A 1852066 unspecified role unspecified role neutral component neutral component transmembrane region sufficient to bind sufficient binding region sufficient to bind range range range range false n-terminal toxr tag tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P02724-PRO_0000012134:p.Gly83Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 2.0 physical association physical association false false false Fig. 2B gp1ba-gp1bb-5 1852062 N/A 1852065 unspecified role unspecified role neutral component neutral component P13224:p.Cys141_Pro145delinsGlySerGlyGlyGly mutation mutation certain certain sequence position certain certain certain sequence position certain false P13224:p.Cys173Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 2.0 N/A 1852067 unspecified role unspecified role neutral component neutral component transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P07359:p.Glu513Gly mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 1852086 1852084 direct interaction direct interaction false false false Fig. 4 Two disulphide bridges form between the alpha and beta peptides gp1ba-gp1bb-2 1852062 N/A 1852068 unspecified role unspecified role neutral component neutral component transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14770:p.Ser142_Trp143delinsGlySer mutation mutation certain certain sequence position certain certain certain sequence position certain false P14770:p.Cys170Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 1852065 unspecified role unspecified role neutral component neutral component P13224:p.Cys141_Pro145delinsGlySerGlyGlyGly mutation mutation certain certain sequence position certain certain certain sequence position certain false P13224:p.Cys173Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 2.0 N/A 1852067 unspecified role unspecified role neutral component neutral component transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P07359:p.Glu513Gly mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 physical association physical association false false false Fig. 4 Two disulphide bridges form between the alpha and beta peptides. Addition of IX-TM significantly facilitated formation of native-like disulfide bonds between alpha and beta peptides in detergent micelles gp1ba-gp1bb-3 1852063 N/A 1852068 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept cys-170 cy5 label cy5 label certain certain sequence position certain certain certain sequence position certain false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14770:p.Ser142_Trp143delinsGlySer mutation mutation certain certain sequence position certain certain certain sequence position certain false P14770:p.Cys170Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 1852065 unspecified role unspecified role neutral component neutral component P13224:p.Cys141_Pro145delinsGlySerGlyGlyGly mutation mutation certain certain sequence position certain certain certain sequence position certain false P13224:p.Cys173Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 1852067 unspecified role unspecified role fluorescence donor fluorescence donor n-terminal 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 n-terminal cy3 label cy3 label n-term range n-terminal range n-term range n-term range n-terminal range n-term range false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P07359:p.Glu513Gly mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Fig. 8B, 8C Two disulphide bridges form between the alpha and beta peptides. Addition of IX-TM significantly facilitated formation of native-like disulfide bonds between alpha and beta peptides in detergent micelles gp1ba-gp1bb-4 1852063 N/A 1852068 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept cys-170 cy5 label cy5 label certain certain sequence position certain certain certain sequence position certain false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14770:p.Ser142_Trp143delinsGlySer mutation mutation certain certain sequence position certain certain certain sequence position certain false P14770:p.Cys170Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 1852068 competitor competitor competitor neutral component neutral component transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P14770:p.Ser142_Trp143delinsGlySer mutation mutation certain certain sequence position certain certain certain sequence position certain false P14770:p.Cys170Asn mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 1852065 unspecified role unspecified role neutral component neutral component P13224:p.Cys141_Pro145delinsGlySerGlyGlyGly mutation mutation certain certain sequence position certain certain certain sequence position certain false P13224:p.Cys173Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 1852067 unspecified role unspecified role fluorescence donor fluorescence donor n-terminal 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 n-terminal cy3 label cy3 label n-term range n-terminal range n-term range n-term range n-terminal range n-term range false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P07359:p.Glu513Gly mutation mutation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Fig. 8B Two disulphide bridges form between the alpha and beta peptides. Addition of IX-TM significantly facilitated formation of native-like disulfide bonds between alpha and beta peptides in detergent micelles gp1bb-2 1852064 N/A 1852065 unspecified role unspecified role neutral component neutral component P13224:p.Cys141_Pro145delinsGlySerGlyGlyGly mutation mutation certain certain sequence position certain certain certain sequence position certain false P13224:p.Cys173Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false transmembrane region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig. 3