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/ Nesprin-1alpha self-associates and binds directly to emerin and lamin A in vitro. Nesprin-1alpha self-associates and binds directly to emerin and lamin A in vitro. 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. 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 Only protein-protein interactions imex curation 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 Accepted 2015-APR-09 by ORCHARD Adult human skeletal muscle library. FEBS letters 2002 imex curation Nesprin-1alpha self-associates and binds directly to emerin and lamin A in vitro. Nesprin-1alpha self-associates and binds directly to emerin and lamin A in vitro. 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. 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 Only protein-protein interactions imex curation 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 Accepted 2015-APR-09 by ORCHARD FEBS letters 2002 imex curation Nesprin-1alpha self-associates and binds directly to emerin and lamin A in vitro. Nesprin-1alpha self-associates and binds directly to emerin and lamin A in vitro. 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. 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 Only protein-protein interactions imex curation 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 Accepted 2015-MAY-05 by ORCHARD FEBS letters 2002 Only protein-protein interactions imex curation imex curation yes-deleted! lmna_human Prelamin-A/C LMNA LMN1 protein protein human Homo sapiens Human METPSQRRATRSGAQASSTPLSPTRITRLQEKEDLQELNDRLAVYIDRVRSLETENAGLRLRITESEEVVSREVSGIKAAYEAELGDARKTLDSVAKERARLQLELSKVREEFKELKARNTKKEGDLIAAQARLKDLEALLNSKEAALSTALSEKRTLEGELHDLRGQVAKLEAALGEAKKQLQDEMLRRVDAENRLQTMKEELDFQKNIYSEELRETKRRHETRLVEIDNGKQREFESRLADALQELRAQHEDQVEQYKKELEKTYSAKLDNARQSAERNSNLVGAAHEELQQSRIRIDSLSAQLSQLQKQLAAKEAKLRDLEDSLARERDTSRRLLAEKEREMAEMRARMQQQLDEYQELLDIKLALDMEIHAYRKLLEGEEERLRLSPSPTSQRSRGRASSHSSQTQGGGSVTKKRKLESTESRSSFSQHARTSGRVAVEEVDEEGKFVRLRNKSNEDQSMGNWQIKRQNGDDPLLTYRFPPKFTLKAGQVVTIWAAGAGATHSPPTDLVWKAQNTWGCGNSLRTALINSTGEEVAMRKLVRSVTVVEDDEDEDGDDLLHHHHGSHCSSSGDPAEYNLRSRTVLCGTCGQPADKASASGSGAQVGGPISSGSSASSVTVTRSYRSVGGSGGGSFGDNLVTRSYLLGNSSPRTQSPQNCSIM E0855F7699F0318B baf_human Barrier-to-autointegration factor BANF1 BAF BCRG1 Breakpoint cluster region protein 1 protein protein human Homo sapiens Human MTTSQKHRDFVAEPMGEKPVGSLAGIGEVLGKKLEERGFDKAYVVLGQFLVLKKDEDLFREWLKDTCGANAKQSRDCFGCLREWCDAFL DIP protein O75531 original sequence version: 123 9A2180A2D284F5D0 emd_human Emerin EDMD STA EMD protein protein human Homo sapiens Human MDNYADLSDTELTTLLRRYNIPHGPVVGSTRRLYEKKIFEYETQRRRLSPPSSSAASSYSFSDLNSTRGDADMYDLPKKEDALLYQSKGYNDDYYEESYFTTRTYGEPESAGPSRAVRQSVTSFPDADAFHHQVHDDDLLSSSEEECKDRERPMYGRDSAYQSITHYRPVSASRSSLDLSYYPTSSSTSFMSSSSSSSSWLTRRAIRPENRAPGAGLGQDRQVPLWGQLLLFLVFVIVLFFIYHFMQAEEGNPF DIP protein P50402 original sequence version: 95 DIP protein P50402 original sequence version: 125 EB62EDD59B7A044F 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. E075A5CDBDAAFC43 syne1-syne1-1 1 7 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 7 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 10 12 physical association physical association (Figure 1) syne1-syne1-2 2 7 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 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 ecoli Escherichia coli (strain K12) 7 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 region 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 18 15 physical association physical association Figure 2 syne1-syne1-3 2 7 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 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 ecoli Escherichia coli (strain K12) 7 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 region 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 25 22 physical association physical association Figure 2-2 banf1-emd-1 2 6 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 ecoli Escherichia coli (strain K12) 5 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 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 in vitro In vitro physical association physical association Figure 3 syne1-lmna-3 2 4 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 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 ecoli Escherichia coli (strain K12) 7 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 region 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 35 38 physical association physical association Figure 4 syne1-emd-1 2 6 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 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 ecoli Escherichia coli (strain K12) 7 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 region 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 45 42 physical association physical association Figure 5B emd-syne1-1 3 6 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 ecoli Escherichia coli (strain K12) 7 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 region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined sr5-7_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro physical association physical association Figure 5D-E nesprin-1alpha SR5-7 binding to Emerin was substoichiometric. emd-syne1-3 3 7 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 sr1-7_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain in vitro In vitro 6 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 ecoli Escherichia coli (strain K12) physical association physical association Figure 5C