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/ hassaine-2001-1 The tyrosine kinase Hck is an inhibitor of HIV-1 replication counteracted by the viral vif protein. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Hassaine G., Courcoul M., Bessou G., Barthalay Y., Picard C., Olive D., Collette Y., Vigne R., Decroly E. Baits were incubated with radiolabeled proteins and interacting proteins were detected after pull-down by SDS-PAGE and autoradiography fig 2, 4, 5 rvigne@inserm-u372.univ-mrs.fr,edecroly@ulb.ac.be J. Biol. Chem. (0021-9258) 2001 hassaine-2001-2 The tyrosine kinase Hck is an inhibitor of HIV-1 replication counteracted by the viral vif protein. human-u937 Homo sapiens histiocytic lymphoma cells u937 Human histiocytic lymphoma cells pull down pull down western blot western blot Immuno blot Immuno blot Hassaine G., Courcoul M., Bessou G., Barthalay Y., Picard C., Olive D., Collette Y., Vigne R., Decroly E. Baits were incubated with radiolabeled proteins and interacting proteins were detected after pull-down by SDS-PAGE and autoradiography fig 2, 4, 5 rvigne@inserm-u372.univ-mrs.fr,edecroly@ulb.ac.be J. Biol. Chem. (0021-9258) 2001 hassaine-2001-3 The tyrosine kinase Hck is an inhibitor of HIV-1 replication counteracted by the viral vif protein. in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Hassaine G., Courcoul M., Bessou G., Barthalay Y., Picard C., Olive D., Collette Y., Vigne R., Decroly E. Baits were incubated with radiolabeled proteins and interacting proteins were detected after pull-down by SDS-PAGE and autoradiography fig 2, 4, 5 rvigne@inserm-u372.univ-mrs.fr,edecroly@ulb.ac.be J. Biol. Chem. (0021-9258) 2001 vif_hv1n5 Virion infectivity factor SOR protein vif protein protein hv1n5 Human immunodeficiency virus type 1 group M subtype B (isolate NY5) (HIV-1) HIV-1 MENRWQVMIVWQVDRMRINTWKRLVKHHMYISRKAKDWFYRHHYESTNPKISSEVHIPLGDAKLVITTYWGLHTGERDWHLGQGVSIEWRKKRYSTQVDPDLADQLIHLHYFDCFSESAIRNTILGRIVSPRCEYQAGHNKVGSLQYLALAALIKPKQIKPPLPSVRKLTEDRWNKPQKTKGHRGSHTMNGH hck_human Tyrosine-protein kinase HCK HCK Hemopoietic cell kinase p59-HCK/p60-HCK Hematopoietic cell kinase p59Hck p61Hck protein protein human Homo sapiens Human MGGRSSCEDPGCPRDEERAPRMGCMKSKFLQVGGNTFSKTETSASPHCPVYVPDPTSTIKPGPNSHNSNTPGIREAGSEDIIVVALYDYEAIHHEDLSFQKGDQMVVLEESGEWWKARSLATRKEGYIPSNYVARVDSLETEEWFFKGISRKDAERQLLAPGNMLGSFMIRDSETTKGSYSLSVRDYDPRQGDTVKHYKIRTLDNGGFYISPRSTFSTLQELVDHYKKGNDGLCQKLSVPCMSSKPQKPWEKDAWEIPRESLKLEKKLGAGQFGEVWMATYNKHTKVAVKTMKPGSMSVEAFLAEANVMKTLQHDKLVKLHAVVTKEPIYIITEFMAKGSLLDFLKSDEGSKQPLPKLIDFSAQIAEGMAFIEQRNYIHRDLRAANILVSASLVCKIADFGLARVIEDNEYTAREGAKFPIKWTAPEAINFGSFTIKSDVWSFGILLMEIVTYGRIPYPGMSNPEVIRALERGYRMPRPENCPEELYNIMMRCWKNRPEERPTFEYIQSVLDDFYTATESQYQQQP khdr1_human KH domain-containing, RNA-binding, signal transduction-associated protein 1 SAM68 Src-associated in mitosis 68 kDa protein KHDRBS1 p21 Ras GTPase-activating protein-associated p62 GAP-associated tyrosine phosphoprotein p62 p68 protein protein human Homo sapiens Human MQRRDDPAARMSRSSGRSGSMDPSGAHPSVRQTPSRQPPLPHRSRGGGGGSRGGARASPATQPPPLLPPSATGPDATVGGPAPTPLLPPSATASVKMEPENKYLPELMAEKDSLDPSFTHAMQLLTAEIEKIQKGDSKKDDEENYLDLFSHKNMKLKERVLIPVKQYPKFNFVGKILGPQGNTIKRLQEETGAKISVLGKGSMRDKAKEEELRKGGDPKYAHLNMDLHVFIEVFGPPCEAYALMAHAMEEVKKFLVPDMMDDICQEQFLELSYLNGVPEPSRGRGVPVRGRGAAPPPPPVPRGRGVGPPRGALVRGTPVRGAITRGATVTRGVPPPPTVRGAPAPRARTAGIQRIPLPPPPAPETYEEYGYDDTYAEQSYEGYEGYYSQSQGDSEYYDYGHGEVQDSYEAYGQDDWNGTRPSLKAPPARPVKGAYREHPYGRY lck_human Tyrosine-protein kinase Lck Protein YT16 Proto-oncogene Lck T cell-specific protein-tyrosine kinase LCK Lymphocyte cell-specific protein-tyrosine kinase p56-LCK Leukocyte C-terminal Src kinase protein protein human Homo sapiens Human MGCGCSSHPEDDWMENIDVCENCHYPIVPLDGKGTLLIRNGSEVRDPLVTYEGSNPPASPLQDNLVIALHSYEPSHDGDLGFEKGEQLRILEQSGEWWKAQSLTTGQEGFIPFNFVAKANSLEPEPWFFKNLSRKDAERQLLAPGNTHGSFLIRESESTAGSFSLSVRDFDQNQGEVVKHYKIRNLDNGGFYISPRITFPGLHELVRHYTNASDGLCTRLSRPCQTQKPQKPWWEDEWEVPRETLKLVERLGAGQFGEVWMGYYNGHTKVAVKSLKQGSMSPDAFLAEANLMKQLQHQRLVRLYAVVTQEPIYIITEYMENGSLVDFLKTPSGIKLTINKLLDMAAQIAEGMAFIEERNYIHRDLRAANILVSDTLSCKIADFGLARLIEDNEYTAREGAKFPIKWTAPEAINYGTFTIKSDVWSFGILLTEIVTHGRIPYPGMTNPEVIQNLERGYRMVRPDNCPEELYQLMRLCWKERPEDRPTFDYLRSVLEDFFTATEGQYQPQP p46108-1 Adapter molecule crk Proto-oncogene c-Crk CRK p38 protein protein human Homo sapiens Human MAGNFDSEERSSWYWGRLSRQEAVALLQGQRHGVFLVRDSSTSPGDYVLSVSENSRVSHYIINSSGPRPPVPPSPAQPPPGVSPSRLRIGDQEFDSLPALLEFYKIHYLDTTTLIEPVSRSRQGSGVILRQEEAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQWWNAEDSEGKRGMIPVPYVEKYRPASASVSALIGGNQEGSHPQPLGGPEPGPYAQPSVNTPLPNLQNGPIYARVIQKRVPNAYDKTALALEVGELVKVTKINVSGQWEGECNGKRGHFPFTHVRLLDQQNPDEDFS grb2_human Growth factor receptor-bound protein 2 GRB2 ASH Adapter protein GRB2 SH2/SH3 adapter GRB2 Protein Ash protein protein human Homo sapiens Human MEAIAKYDFKATADDELSFKRGDILKVLNEECDQNWYKAELNGKDGFIPKNYIEMKPHPWFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLWVVKFNSLNELVDYHRSTSVSRNQQIFLRDIEQVPQQPTYVQALFDFDPQEDGELGFRRGDFIHVMDNSDPNWWKGACHGQTGMFPRNYVTPVNRNV hck-vif-1 In vitro pull-down: bait= hck SH3 domain, prey = HIV-1 vif 297063 N/A 297066 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 in vitro In vitro N/A 297067 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P08631:p.Trp114Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P08631:p.Tyr87Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P08631:p.Tyr89Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P08631:p.Asp96Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false HIV-1 pNL4.3 corresponding to HIV M19921 (HV1N5) was used to amplify vif. Vif binding to GST-SH3(Hck) was enhanced at 350 mM NaCl compared to 150 mM concentration, suggesting that hydrophobic residues participated to the interaction. All 3 parts (N-terminus, central, C-terminus) of Vif seem to contribute to the interaction with hck SH3 with a weaker involvement of the N-terminus. PPLP motif mutation to AALA in Vif decreased the binding when using the 1/3 c-terminal part of Vif but has no effect in context of the full-length. hck-sam68-1 In vitro pull-down: bait= hck SH3 domain, prey = sam68 297063 N/A 297067 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false sh3 domain 76-147 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 297068 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 in vitro In vitro physical association physical association false false false lck-sam68-1 In vitro pull-down: bait= Lck SH3 domain, prey = sam68 297063 N/A 297069 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false sh3 domain sufficient to bind sufficient binding region sufficient to bind range range range range false ecoli Escherichia coli (strain K12) N/A 297068 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 in vitro In vitro physical association physical association false false false crk-sam68-1 In vitro pull-down: bait= CRK II SH3 domains, prey = sam68 297063 N/A 297070 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false sh3 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 297068 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 in vitro In vitro physical association physical association false false false grb2-sam68-1 In vitro pull-down: bait= Grb2 SH3 domain 1, prey = sam68 297063 N/A 297068 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 in vitro In vitro N/A 297071 unspecified role unspecified role bait bait sh3 domain 1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false grb2-sam68-2 In vitro pull-down: bait= Grb2 SH3 domain 2, prey = sam68 297063 N/A 297068 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 in vitro In vitro N/A 297071 unspecified role unspecified role bait bait sh3 domain 2 sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false hck-vif-2 In vivo pull-down: bait= hck SH3 domain, prey = HIV-1 vif 297064 N/A 297066 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 297067 unspecified role unspecified role prey prey physical association physical association false false false HIV-1 pNL4.3 corresponding to HIV M19921 (HV1N5) was used to amplify vif. hck-vif-3 Vif inhibits hck autophosphorylation 297065 N/A 297067 self self unspecified role unspecified role human-u937 Homo sapiens histiocytic lymphoma cells u937 Human histiocytic lymphoma cells N/A 297066 inhibitor inhibitor unspecified role unspecified role region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false HIV-1 pNL4.3 corresponding to HIV M19921 (HV1N5) was used to amplify vif. vif also inhibits hck phosphorylation of enolase in vitro.