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/ mislow-2002-2 in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM. Cardiac - Interactions involving cardiac related proteins Emerin and BAF constructs have been entered as of Human origin. However the quoted references (nos 16 and 27) only describe BAF in the pET15b vector, not Emerin; and the earlier 1998 reference (no 27) desribes Murine BAF. Only the later publication -Reference 16 - describes In Vitro translated Human BAF, so this has been taken as the species of origin for BAF. Emerin construct details have been taken from ref16- Lee-2001; which refers to the expression vector pET11c with an N-terminal T7 tag. Nesprin-1alpha (GenBank accession number AAL38031) has been mapped to Uniprot identifier Q8NF91-11 (isoform 11, 1139 amino acids). emcnally@medicine.bsd.uchicago.edu,elizabeth.mcnally@northwestern.edu emcnally@medicine.bsd.uchicago.edu FEBS letters 2002 imex curation Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM., Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM. Only protein-protein interactions imex curation mislow-2002-3 in vitro In vitro solid phase assay solid phase assay predetermined predetermined participant predetermined Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM., Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM. Cardiac - Interactions involving cardiac related proteins Emerin was immobilized in microtiter wells, washed, and incubated with soluble 35S-labeled nesprin-1alpha fragments. Nesprin-1alpha (GenBank accession number AAL38031) has been mapped to Uniprot identifier Q8NF91-11 (isoform 11, 1139 amino acids). emcnally@medicine.bsd.uchicago.edu,elizabeth.mcnally@northwestern.edu emcnally@medicine.bsd.uchicago.edu FEBS letters 2002 Only protein-protein interactions imex curation imex curation mislow-2002-1 yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM. Cardiac - Interactions involving cardiac related proteins Nesprin-1alpha (GenBank accession number AAL38031) has been mapped to Uniprot identifier Q8NF91-11 (isoform 11, 1139 amino acids). emcnally@medicine.bsd.uchicago.edu,elizabeth.mcnally@northwestern.edu emcnally@medicine.bsd.uchicago.edu Adult human skeletal muscle library. FEBS letters 2002 imex curation Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM., Mislow JM., Holaska JM., Kim MS., Lee KK., Segura-Totten M., Wilson KL., McNally EM. Only protein-protein interactions imex curation q8nf91-11 Nesprin-1 Enaptin Myocyte nuclear envelope protein 1 Nuclear envelope spectrin repeat protein 1 Synaptic nuclear envelope protein 1 131kDa myne-1 SYNE1 C6orf98 KIAA0796 KIAA1262 KIAA1756 MYNE1 KASH domain-containing protein 1 protein protein human Homo sapiens Human MRLEEQKKKLAFLLKDWEKCEKGIADSLEKLRTFKKKLSQSLPDHHEELHAEQMRCKELENAVGSWTDDLTQLSLLKDTLSAYISADDISILNERVELLQRQWEELCHQLSLRRQQIGERLNEWAVFSEKNKELCEWLTQMESKVSQNGDILIEEMIEKLKKDYQEEIAIAQENKIQLQQMGERLAKASHESKASEIEYKLGKVNDRWQHLLDLIAARVKKLKETLVAVQQLDKNMSSLRTWLAHIESELAKPIVYDSCNSEEIQRKLNEQQELQRDIEKHSTGVASVLNLCEVLLHDCDACATDAECDSIQQATRNLDRRWRNICAMSMERRLKIEETWRLWQKFLDDYSRFEDWLKSSERTAAFPSSSGVIYTVAKEELKKFEAFQRQVHECLTQLELINKQYRRLARENRTDSACSLKQMVHEGNQRWDNLQKRVTSILRRLKHFIGQREEFETARDSILVWLTEMDLQLTNIEHFSECDVQAKIKQLKAFQQEISLNHNKIEQIIAQGEQLIEKSEPLDAAIIEEELDELRRYCQEVFGRVERYHKKLIRLPLPDDEHDLSDRELELEDSAALSDLHWHDRSADSLLSPQPSSNLSLSLAQPLRSERSGRDTPASVDSIPLEWDHDYDLSRDLESAMSRALPSEDEEGQDDKDFYLRGAVGLSGDHSALESQIRQLGKALDDSRFQIQQTENIIRSKTPTGPELDTSYKGYMKLLGECSSSIDSVKRLEHKLKEEEESLPGFVNLHSTETQTAGVIDRWELLQAQALSKELRMKQNLQKWQQFNSDLNSIWAWLGDTEEELEQLQRLELSTDIQTIELQIKKLKELQKAVDHRKAIILSINLCSPEFTQADSKESRDLQDRLSQMNGRWDRVCSLLEEWRGLLQDALMQCQGFHEMSHGLLLMLENIDRRKNEIVPIDSNLDAEILQDHHKQLMQIKHELLESQLRVASLQDMSCQLLVNAEGTDCLEAKEKVHVIGNRLKLLLKEVSRHIKELEKLLDVSSSQQDLSSWSSADELDTSGSVSPTSGRSTPNRQKTPRGKCSLSQPGPSVSSPHSRSTKGGSDSSLSEPGPGRSGRGFLFRVLRAALPLQLLLLLLIGLACLVPMSEEDYSCALSNNFARSFHPMLRYTNGPPPL Muscle-specific. emd_human Emerin EDMD STA EMD protein protein human Homo sapiens Human MDNYADLSDTELTTLLRRYNIPHGPVVGSTRRLYEKKIFEYETQRRRLSPPSSSAASSYSFSDLNSTRGDADMYDLPKKEDALLYQSKGYNDDYYEESYFTTRTYGEPESAGPSRAVRQSVTSFPDADAFHHQVHDDDLLSSSEEECKDRERPMYGRDSAYQSITHYRPVSASRSSLDLSYYPTSSSTSFMSSSSSSSSWLTRRAIRPENRAPGAGLGQDRQVPLWGQLLLFLVFVIVLFFIYHFMQAEEGNPF baf_human Barrier-to-autointegration factor BANF1 BAF BCRG1 Breakpoint cluster region protein 1 protein protein human Homo sapiens Human MTTSQKHRDFVAEPMGEKPVGSLAGIGEVLGKKLEERGFDKAYVVLGQFLVLKKDEDLFREWLKDTCGANAKQSRDCFGCLREWCDAFL lmna_human Prelamin-A/C LMNA LMN1 protein protein human Homo sapiens Human METPSQRRATRSGAQASSTPLSPTRITRLQEKEDLQELNDRLAVYIDRVRSLETENAGLRLRITESEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVREEFKELKARNTKKEGDLIAAQARLKDLEALLNSKEAALSTALSEKRTLEGELHDLRGQVAKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETRLVEIDNGKQREFESRLADALQELRAQHEDQVEQYKKELEKTYSAKLDNARQSAERNSNLVGAAHEELQQSRIRIDSLSAQLSQLQKQLAAKEAKLRDLEDSLARERDTSRRLLAEKEREMAEMRARMQQQLDEYQELLDIKLALDMEIHAYRKLLEGEEERLRLSPSPTSQRSRGRASSHSSQTQGGGSVTKKRKLESTESRSSFSQHARTSGRVAVEEVDEEGKFVRLRNKSNEDQSMGNWQIKRQNGDDPLLTYRFPPKFTLKAGQVVTIWAAGAGATHSPPTDLVWKAQNTWGCGNSLRTALINSTGEEVAMRKLVRSVTVVEDDEDEDGDDLLHHHHGSHCSSSGDPAEYNLRSRTVLCGTCGQPADKASASGSGAQVGGPISSGSSASSVTVTRSYRSVGGSGGGSFGDNLVTRSYLLGNSSPRTQSPQNCSIM syne1-syne1-2 1791124 N/A 1791127 unspecified role unspecified role prey prey sr_5-6_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 1791127 unspecified role unspecified role bait bait sr3_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 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 1791136 1791133 physical association physical association false false false Figure 2 syne1-syne1-3 1791124 N/A 1791127 unspecified role unspecified role prey prey sr_2-3_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 1791127 unspecified role unspecified role bait bait sr5_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 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 1791143 1791140 physical association physical association false false false Figure 2-2 banf1-emd-1 1791124 N/A 1791128 unspecified role unspecified role prey prey n_terminus t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 1791129 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 false n_terminus 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 in vitro In vitro physical association physical association false false false Figure 3 syne1-lmna-3 1791124 N/A 1791130 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain undetermined undetermined sequence position undetermined false n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 1791127 unspecified role unspecified role bait bait sr5-7_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 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 1791156 1791153 physical association physical association false false false Figure 4 syne1-emd-1 1791124 N/A 1791128 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n_terminus t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) N/A 1791127 unspecified role unspecified role bait bait sr1-5_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 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 1791163 1791160 physical association physical association false false false Figure 5B emd-syne1-1 1791125 N/A 1791128 unspecified role unspecified role bait bait n_terminus t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 1791127 unspecified role unspecified role prey prey sr1-5_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false sr5-7_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 1.0 physical association physical association false false false Figure 5D-E nesprin-1alpha SR5-7 binding to Emerin was substoichiometric. emd-syne1-3 1791125 N/A 1791127 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 false sr1-7_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 2.0 N/A 1791128 unspecified role unspecified role bait bait n_terminus t7 tag t7 tag MASMTGGQQMG epitope tag MASMTGGQQMG epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 physical association physical association false false false 1791125 Figure 5C syne1-syne1-1 1791126 N/A 1791127 unspecified role unspecified role bait bait sr_5-6_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 1791127 unspecified role unspecified role prey prey sr_2-3_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1791182 1791180 physical association physical association false false false (Figure 1)