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/ Subunits of the Saccharomyces cerevisiae signal recognition particle required for its functional expression. Subunits of the Saccharomyces cerevisiae signal recognition particle required for its functional expression. EMBO J. (0261-4189) 1994 imex curation Brown JD., Hann BC., Medzihradszky KF., Niwa M., Burlingame AL., Walter P. Immunoaffinity chromatography with anti-Sec65p antibodies was carried out on yeast post-ribosomal supernatant fractionated by DEAE-Sepaharose chromatography. Matter eluted from the column was precipitated in TCA and subjected to SDS-PAGE. Bands were excised from gels and proteins digested in situ with trypsin and their mass determined by electrospray mass spectrometry. Peptides were then sequenced by either Edman degradation or tandem mass spectrometry. Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Brown JD., Hann BC., Medzihradszky KF., Niwa M., Burlingame AL., Walter P. Immunoaffinity chromatography with anti-Sec65p antibodies was carried out on yeast post-ribosomal supernatant fractionated by DEAE-Sepaharose chromatography. Matter eluted from the column was precipitated in TCA and subjected to SDS-PAGE. Bands were excised from gels and proteins digested in situ with trypsin and their mass determined by electrospray mass spectrometry. Peptides were then sequenced by either Edman degradation or tandem mass spectrometry. Accepted 2007-JUN-17 by JYOTI. EMBO J. (0261-4189) 1994 Only protein-protein interactions imex curation Subunits of the Saccharomyces cerevisiae signal recognition particle required for its functional expression. Subunits of the Saccharomyces cerevisiae signal recognition particle required for its functional expression. EMBO J. (0261-4189) 1994 imex curation Brown JD., Hann BC., Medzihradszky KF., Niwa M., Burlingame AL., Walter P. Immunoaffinity chromatography with anti-Sec65p antibodies was carried out on yeast post-ribosomal supernatant fractionated by DEAE-Sepaharose chromatography. Matter eluted from the column was precipitated in TCA and subjected to SDS-PAGE. Bands were excised from gels and proteins digested in situ with trypsin and their mass determined by electrospray mass spectrometry. Peptides were then sequenced by either Edman degradation or tandem mass spectrometry. Only protein-protein interactions in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom weight by comassie molecular weight estimation by coomasie staining weight by comassie Brown JD., Hann BC., Medzihradszky KF., Niwa M., Burlingame AL., Walter P. Immunoaffinity chromatography with anti-Sec65p antibodies was carried out on yeast post-ribosomal supernatant fractionated by DEAE-Sepaharose chromatography. Matter eluted from the column was precipitated in TCA and subjected to SDS-PAGE. Bands were excised from gels and proteins digested in situ with trypsin and their mass determined by electrospray mass spectrometry. Peptides were then sequenced by either Edman degradation or tandem mass spectrometry. Accepted 2007-JUN-17 by JYOTI. EMBO J. (0261-4189) 1994 Only protein-protein interactions imex curation Subunits of the Saccharomyces cerevisiae signal recognition particle required for its functional expression. Subunits of the Saccharomyces cerevisiae signal recognition particle required for its functional expression. EMBO J. (0261-4189) 1994 imex curation Brown JD., Hann BC., Medzihradszky KF., Niwa M., Burlingame AL., Walter P. Immunoaffinity chromatography with anti-Sec65p antibodies was carried out on yeast post-ribosomal supernatant fractionated by DEAE-Sepaharose chromatography. Matter eluted from the column was precipitated in TCA and subjected to SDS-PAGE. Bands were excised from gels and proteins digested in situ with trypsin and their mass determined by electrospray mass spectrometry. Peptides were then sequenced by either Edman degradation or tandem mass spectrometry. Only protein-protein interactions in vitro In vitro affinity chrom affinity chromatography technology Affinity purification Affinity purification affinity chrom ms participant Identification by mass spectrometry ms participant Brown JD., Hann BC., Medzihradszky KF., Niwa M., Burlingame AL., Walter P. Immunoaffinity chromatography with anti-Sec65p antibodies was carried out on yeast post-ribosomal supernatant fractionated by DEAE-Sepaharose chromatography. Matter eluted from the column was precipitated in TCA and subjected to SDS-PAGE. Bands were excised from gels and proteins digested in situ with trypsin and their mass determined by electrospray mass spectrometry. Peptides were then sequenced by either Edman degradation or tandem mass spectrometry. Accepted 2007-JUN-17 by JYOTI. EMBO J. (0261-4189) 1994 Only protein-protein interactions imex curation srp72_yeast Signal recognition particle subunit SRP72 YPL210C SRP72 Signal recognition particle 72 kDa protein homolog protein protein yeast Saccharomyces cerevisiae Baker's yeast MAKDNLTNLLSQLNIQLSQDEHSQVEQTCVKLLDSGCENPADVFRRCLVAVIQQDKYQKALHYLKKFKHIDDKYGRKFALEKLYIFYKLNMPDEFNTLYTAIITDDLDTVLKKDIESLRGILHVRAQYCYKNGLYQEAFKIYQHLASHNEKDQDSQIELSCNERVPLSVATELMNRSPLVTPMDESSYDLLFNESFIMASVGKYDKAIELLEKALQGATNEGYQNDINTIKLQLSFVLQMVGKTAQSKEILKGLLQELKADSPFSLICQNNLNAFVDFSKYNTNFNLLLRELNVEKLNTFNLQTFTHEQWSNIQRNVLFLRLFNNVKIHSQESLLSRTFDKYSKLVDNVTLESYKTQAKKLYHHTTKTILSGTDGSTIGILLLTIQLLIIEKEWENAIRIGELFLNESWKSSFEKFNDSQAIVCYILFELYKIKGRNNSKSVLLKKLGSVRVQLSGKIQENIPFWKHVGFELLSMGNAKESKALLREISNFSKGDADVLVDRVVSSDSLDIAQGIDLVRDIDIDKLIQLGVKPLESSAKRSKNTAVSKVQKRKVLELKKKRKIKRLEKFLQGRDTSKLPDPERWLPLRDRSTYRPKKKQQGAKQTQGGAMNKKSEQALDISKKGKPTVNKKPKNKKKGRK DB2F536212F0871B srp14_yeast Signal recognition particle subunit SRP14 SRP14 YDL092W D2404 Signal recognition particle 14 kDa protein homolog protein protein yeast Saccharomyces cerevisiae Baker's yeast MANTGCLSPGAFLSKVPEFFQTANEKHITVRLTAKRLIEHDPVEGNLEFDSTNHPDYDVSKKASEISVSSRSDREYPLLIRMSYGSHDKKTKCSTVVKASELDQFWQEYSSVFKGGMQNLIKKKKKKSKNGTISKTGKKNKVAKKN 4AF76A26018367EF srp21_yeast Signal recognition particle subunit SRP21 SRP21 YKL122C YKL527 Signal recognition particle 21 kDa protein protein protein yeast Saccharomyces cerevisiae Baker's yeast MSVKPIDNYITNSVRLFEVNPSQTLFSISYKPPTQKTDTKVSFRTHNSHLSLNYKFTTNKSKDVSRLLSALGPRGVSITPGKIEKIAQSKKKNNKIKESSKKIKGKSIQDIVGLATLIVNTDVEKSDPAAKKTATEPKQKANAVQNNNGNSAASKKKKNKNKGKKKR 19C85378074C6F46 srp68_yeast Signal recognition particle subunit SRP68 YPL243W SRP68 Signal recognition particle 68 kDa protein homolog protein protein yeast Saccharomyces cerevisiae Baker's yeast MVAYSPIIATYGNRAEQFLETDSDFAKYHAKLNKKLQHLRSRCHLVTKDTKKYSSKNKYGEINSEDYDNKTKLIGVLILLHAERDLALAETLKLRARQRGKLKKSEEKVLSTRLKKACKTADKLVNVTQNEQQWITRAQYLAFAKLVHSEYLINGKRFKRKDNAKISNNLALVFAALEHLKNLSLLAEEVVDNIVNKYQYSLKQYAGNLITTPEINNFIVERVQSDENKDDELVKLLLDNGFNMKKITTSTEDQKVTTNINWRSFNAKIIDAEVAQFLEQGLSIHPTQITQYTQRLSKLEKALDRHEFFIANHDDQDDIDEMVENSSENNQIILAYIKYNILLTSISRERDLFTHLWNQWLKLNTSLPSKLTKYKEMERIVKNLTKYLSDIMELPGVYSDDELLSQLDLCKLYFQLFLNTGCLSVLYQSKGRYMEALALYVDAYRRLENKLSEIESLDEILLPANLLSLNSVRSLQKRIENGGNSVITLAEYEKRNHGGSLGKYDLTVIEKLDSKKILPTDIQLKNLFPLKPKMLPIPSKPTLFDLAFNYITYDKQEPSASQVKDSVTETESISQTPISNEQTEGEPKKKRGFLGLFGR 241749187CE95499 srp54_yeast Signal recognition particle subunit SRP54 SRH1 P9513.14 YPR088C SRP54 Signal recognition particle 54 kDa protein homolog protein protein yeast Saccharomyces cerevisiae Baker's yeast MVLADLGKRINSAVNNAISNTQDDFTTSVDVMLKGIVTALLESDVNIALVSKLRNNIRSQLLSENRSEKSTTNAQTKKLIQKTVFDELCKLVTCEGSEEKAFVPKKRKTNIIMFVGLQGSGKTTSCTKLAVYYSKRGFKVGLVCADTFRAGAFDQLKQNAIRARIPFYGSYTETDPAKVAEEGINKFKKEKFDIIIVDTSGRHHQEEELFQEMIEISNVIKPNQTIMVLDASIGQAAEQQSKAFKESSDFGAIILTKMDGHARGGGAISAVAATNTPIIFIGTGEHIHDLEKFSPKSFISKLLGIGDIESLFEQLQTVSNKEDAKATMENIQKGKFTLLDFKKQMQTIMKMGPLSNIAQMIPGMSNMMNQVGEEETSQKMKKMVYVLDSMTKEELESDGRMFIEEPTRMVRVAKGSGTSVFEVEMILMQQQMMARMAQTATQQQPGAPGANARMPGMPNMPGMPNMPGMPNMPGMPKVTPQMMQQAQQKLKQNPGLMQNMMNMFGGGMGGGMGGGMPDMNEMMKMMQDPQMQQMAKQFGMG 05B1B2C262932F17 05B1B2C262932F17 sec65_yeast Signal recognition particle subunit SEC65 YM8339.14C YML105C SEC65 protein protein yeast Saccharomyces cerevisiae Baker's yeast MPRLEEIDDFNDIDDLDMELAELDPSLRTPIAPKITPKVVRSQDQENPAFLPGTNNNSNSNNNSSNEKEQLSFINPKTGKVERSEAISKKDLEEVKRFQVLYPCYFDINRSHKEGRRVPKELAVENPLAKTMADAVRELGILCIFEGEKCHPQDFGNPGRIRVLFKENGQLIGAATKFKGGKRQLMKAVGEYMKRHPTTIESLREIPYGPDFDNIEFKKIPRVKGFKMNEIVPLHSPFLMGHPMTKSVYETPKITAAEKSFKPPKNKYKVVRR 4F4A6CBB9225E385 sec65-srp14-1 1 9 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 1 B In addition to Sec65p and Srp54p proteins (identified by comigration of bands in non-native immunoprecipitations), novel species of 19 and 66 kDa were immunoprecipitated from 16S fractions of a gradient through which wild type cell extract was sedimented As will be shown below, the 66 kDa band is a doublet corresponding to yeast Srp68p and Srp72p subunits, and the 19 kDa band corresponds to the yeast Srpl4p homolog. The doublet of bands migrating at 66 kDA consists of Srp68p and Srp72p. These two proteins were not resolved in the gel and thus have not been added to this interaction. sec65-srp14-2 2 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey association association The doublet of bands migrating at 66 kDA consists of Srp68p and Srp72p. These two proteins were not resolved in the gel and thus have not been added to this interaction. 2 A The final fraction (lane 8) consisted of six proteins in approximately equimolar amounts: a doublet of bands migrating at 66 kDa (Srp68p and Srp72p) and single bands at 60 (Srp54p), 34 (Sec65p), 21 (Srp2lp) and 19 kDa (Srpl4p). The identification of an additional protein, Srp2lp, in this purification, not seen in the immunoprecipitations of [35S]methionine-labeled proteins, suggested that it lacked methionine residues and was therefore not visible in Figure lB. sec65-srp14-3 3 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role prey prey 4 unspecified role unspecified role prey prey association association Table 1