DIP Database of Interacting Proteins 611 Young Drive East; Los Angeles CA 90095; USA http://dip.doe-mbi.ucla.edu/ dip@mbi.ucla.edu Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Science (New York, N.Y.) 2000 imex curation Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro affinity techniques affinity technology affinity techniques weight by staining molecular weight estimation by staining weight by staining Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. Purified Smad-binding fragment from SARA and the MH2 domain from Smad2 (residues 241-467) mixed and incubated. The complex was then passed through a cation-exchange column to which Smad2 MH2 domain binds avidly. GST-SARA SBD in isolation does not bind this column. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu Accepted 2020-AUG-10 AT 14:35 BST AT 14:35 BST by MGT Science (New York, N.Y.) 2000 Only protein-protein interactions imex curation imex curation Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Science (New York, N.Y.) 2000 imex curation Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu Accepted 2020-AUG-10 AT 14:35 BST AT 14:35 BST by MGT Science (New York, N.Y.) 2000 Only protein-protein interactions imex curation imex curation Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Science (New York, N.Y.) 2000 imex curation Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 confocal microscopy confocal microscopy anti tag immunost anti tag immunostaining anti tag immunost Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu Accepted 2020-AUG-10 AT 14:35 BST AT 14:35 BST by MGT Science (New York, N.Y.) 2000 Only protein-protein interactions imex curation imex curation Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Structural basis of Smad2 recognition by the Smad anchor for receptor activation. Science (New York, N.Y.) 2000 imex curation Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu 28-Sep-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Wu G., Chen YG., Ozdamar B., Gyuricza CA., Chong PA., Wrana JL., Massagué J., Shi Y. The current version of SARA in Uniprot (2020) has an extra 102 residues at the N-terminus as compared to the sequence in 2000. yshi@molbio.princeton.edu Accepted 2020-AUG-10 AT 14:35 BST AT 14:35 BST by MGT Science (New York, N.Y.) 2000 Only protein-protein interactions imex curation imex curation zfyv9_human Zinc finger FYVE domain-containing protein 9 ZFYVE9 MADHIP SARA SMADIP Mothers against decapentaplegic homolog-interacting protein Smad anchor for receptor activation Receptor activation anchor Novel serine protease protein protein human Homo sapiens Human MENYFQAEAYNLDKVLDEFEQNEDETVSSTLLDTKWNKILDPPSHRLSFNPTLASVNESAVSNESQPQLKVFSLAHSAPLTTEEEDHCANGQDCNLNPEIATMWIDENAVAEDQLIKRNYSWDDQCSAVEVGEKKCGNLACLPDEKNVLVVAVMHNCDKRTLQNDLQDCNNYNSQSLMDAFSCSLDNENRQTDQFSFSINESTEKDMNSEKQMDPLNRPKTEGRSVNHLCPTSSDSLASVCSPSQLKDDGSIGRDPSMSAITSLTVDSVISSQGTDGCPAVKKQENYIPDEDLTGKISSPRTDLGSPNSFSHMSEGILMKKEPAEESTTEESLRSGLPLLLKPDMPNGSGRNNDCERCSDCLVPNEVRADENEGYEHEETLGTTEFLNMTEHFSESQDMTNWKLTKLNEMNDSQVNEEKEKFLQISQPEDTNGDSGGQCVGLADAGLDLKGTCISESEECDFSTVIDTPAANYLSNGCDSYGMQDPGVSFVPKTLPSKEDSVTEEKEIEESKSECYSNIYEQRGNEATEGSGLLLNSTGDLMKKNYLHNFCSQVPSVLGQSSPKVVASLPSISVPFGGARPKQPSNLKLQIPKPLSDHLQNDFPANSGNNTKNKNDILGKAKLGENSATNVCSPSLGNISNVDTNGEHLESYEAEISTRPCLALAPDSPDNDLRAGQFGISARKPFTTLGEVAPVWVPDSQAPNCMKCEARFTFTKRRHHCRACGKVFCASCCSLKCKLLYMDRKEARVCVICHSVLMNAQAWENMMSASSQSPNPNNPAEYCSTIPPLQQAQASGALSSPPPTVMVPVGVLKHPGAEVAQPREQRRVWFADGILPNGEVADAAKLTMNGTSSAGTLAVSHDPVKPVTTSPLPAETDICLFSGSITQVGSPVGSAMNLIPEDGLPPILISTGVKGDYAVEEKPSQISVMQQLEDGGPDPLVFVLNANLLSMVKIVNYVNRKCWCFTTKGMHAVGQSEIVILLQCLPDEKCLPKDIFNHFVQLYRDALAGNVVSNLGHSFFSQSFLGSKEHGGFLYVTSTYQSLQDLVLPTPPYLFGILIQKWETPWAKVFPIRLMLRLGAEYRLYPCPLFSVRFRKPLFGETGHTIMNLLADFRNYQYTLPVVQGLVVDMEVRKTSIKIPSNRYNEMMKAMNKSNEHVLAGGACFNEKADSHLVCVQNDDGNYQTQAISIHNQPRKVTGASFFVFSGALKSSSGYLAKSSIVEDGVMVQITAENMDSLRQALREMKDFTITCGKADAEEPQEHIHIQWVDDDKNVSKGVVSPIDGKSMETITNVKIFHGSEYKANGKVIRWTEVFFLENDDQHNCLSDPADHSRLTEHVAKAFCLALCPHLKLLKEDGMTKLGLRVTLDSDQVGYQAGSNGQPLPSQYMNDLDSALVPVIHGGACQLSEGPVVMELIFYILENIV ADE5CB530C1272C6 smad2_human Mothers against decapentaplegic homolog 2 SMAD2 MADH2 MADR2 Mad-related protein 2 SMAD family member 2 JV18-1 protein protein human Homo sapiens Human MSSILPFTPPVVKRLLGWKKSAGGSGGAGGGEQNGQEEKWCEKAVKSLVKKLKKTGRLDELEKAITTQNCNTKCVTIPSTCSEIWGLSTPNTIDQWDTTGLYSFSEQTRSLDGRLQVSHRKGLPHVIYCRLWRWPDLHSHHELKAIENCEYAFNLKKDEVCVNPYHYQRVETPVLPPVLVPRHTEILTELPPLDDYTHSIPENTNFPAGIEPQSNYIPETPPPGYISEDGETSDQQLNQSMDTGSPAELSPTTLSPVNHSLDLQPVTYSEPAFWCSIAYYELNQRVGETFHASQPSLTVDGFTDPSNSERFCLGLLSNVNRNATVEMTRRHIGRGVRLYYIGGEVFAECLSDSAIFVQSPNCNQRYGWHPATVCKIPPGCNLKIFNNQEFAALLAQSVNQGFEAVYQLTRMCTIRMSFVKGWGAEYRRQTVTSTPCWIELHLNGPLQWLDKVLTQMGSPSVRCSSMS DIP protein Q15796 original sequence version: 174 DIP protein Q15796 original sequence version: 100 DIP protein Q15796-1 original sequence version: 141 95406DB5FC0AA4C9 smad2-zfyve9-1 4 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed purified purified MH2 domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed purified purified sbd domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Table 1 smad2-zfyve9-2 2 6 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed Q15796:p.Ser308_Glu309delinsAsnLys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain SE NK Wed Sep 16 09:48:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'Q15796:p.Ser308_Glu309delins AsnLys' Q15796:p.Gln294Ser mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Q S Q15796:p.Arg337His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R H Q15796:p.Asn381Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain N S FLAG tag FLAG tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined MH2 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed MYC tag MYC tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig 1A smad2-zfyve9-3 1 6 unspecified role unspecified role bait bait purified purified 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 MH2 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey purified purified 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 sbd domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain O95405:p.Tyr782Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y A O95405:p.Tyr782Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y E O95405:p.Cys783Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain C A O95405:p.Cys783Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain C E O95405:p.Pro788Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain P A O95405:p.Pro788Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain P E O95405:p.Gln790Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Q A O95405:p.Gln793Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain Q A O95405:p.Val805Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V A O95405:p.Val805Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V E ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 1C zfyve9-smad2-2 3 6 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed Q15796:p.Asn381Ser mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain N S FLAG tag FLAG tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed MYC tag MYC tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain colocalization colocalization 4B