MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. EMBO J. (0261-4189) 2011 imex curation Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. EMBO J. (0261-4189) 2011 imex curation Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. Only protein-protein interactions in vitro In vitro bn-page blue native page bn-page predetermined predetermined participant predetermined Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. EMBO J. (0261-4189) 2011 imex curation Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. Only protein-protein interactions in vitro In vitro ion exchange chrom ion exchange chromatography IEC IEC ion exchange chrom predetermined predetermined participant predetermined Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. TGF-β signalling is mediated by two autonomously functioning TβRI:TβRII pairs. EMBO J. (0261-4189) 2011 imex curation Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. Only protein-protein interactions in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Huang T., David L., Mendoza V., Yang Y., Villarreal M., De K., Sun L., Fang X., López-Casillas F., Wrana JL., Hinck AP. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation tgfr1_human TGF-beta receptor type-1 TGFBR1 Transforming growth factor-beta receptor type I TGF-beta type I receptor Serine/threonine-protein kinase receptor R4 Activin receptor-like kinase 5 ALK5 SKR4 Activin A receptor type II-like protein kinase of 53kD protein protein human Homo sapiens Human MEAAVAAPRPRLLLLVLAAAAAAAAALLPGATALQCFCHLCTKDNFTCVTDGLCFVSVTETTDKVIHNSMCIAEIDLIPRDRPFVCAPSSKTGSVTTTYCCNQDHCNKIELPTTVKSSPGLGPVELAAVIAGPVCFVCISLMLMVYICHNRTVIHHRVPNEEDPSLDRPFISEGTTLKDLIYDMTTSGSGSGLPLLVQRTIARTIVLQESIGKGRFGEVWRGKWRGEEVAVKIFSSREERSWFREAEIYQTVMLRHENILGFIAADNKDNGTWTQLWLVSDYHEHGSLFDYLNRYTVTVEGMIKLALSTASGLAHLHMEIVGTQGKPAIAHRDLKSKNILVKKNGTCCIADLGLAVRHDSATDTIDIAPNHRVGTKRYMAPEVLDDSINMKHFESFKRADIYAMGLVFWEIARRCSIGGIHEDYQLPYYDLVPSDPSVEEMRKVVCEQKLRPNIPNRWQSCEALRVMAKIMRECWYANGAARLTALRIKKTLSQLSQQEGIKM DIP protein P36897 original sequence version: 161 179F11404725DDCB tgfr2_human TGF-beta receptor type-2 TGFBR2 Transforming growth factor-beta receptor type II TGF-beta type II receptor protein protein human Homo sapiens Human MGRGLLRGLWPLHIVLWTRIASTIPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFSTCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASPKCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPDLLLVIFQVTGISLLPPLGVAISVIIIFYCYRVNRQQKLSSTWETGKTRKLMEFSEHCAIILEDDRSDISSTCANNINHNTELLPIELDTLVGKGRFAEVYKAKLKQNTSEQFETVAVKIFPYEEYASWKTEKDIFSDINLKHENILQFLTAEERKTELGKQYWLITAFHAKGNLQEYLTRHVISWEDLRKLGSSLARGIAHLHSDHTPCGRPKMPIVHRDLKSSNILVKNDLTCCLCDFGLSLRLDPTLSVDDLANSGQVGTARYMAPEVLESRMNLENVESFKQTDVYSMALVLWEMTSRCNAVGEVKDYEPPFGSKVREHPCVESMKDNVLRDRGRPEIPSFWLNHQGIQMVCETLTECWDHDPEARLTAQCVAERFSELEHLDRLSGRSCSEEKIPEDGSLNTTK DIP protein P37173 original sequence version: 163 C541DA751FFBDBEB tgfb3_human Transforming growth factor beta-3 proprotein TGFB3 protein protein human Homo sapiens Human MKMHLQRALVVLALLNFATVSLSLSTCTTLDFGHIKKKRVEAIRGQILSKLRLTSPPEPTVMTHVPYQVLALYNSTRELLEEMHGEREEGCTQENTESEYYAKEIHKFDMIQGLAEHNELAVCPKGITSKVFRFNVSSVEKNRTNLFRAEFRVLRVPNPSSKRNEQRIELFQILRPDEHIAKQRYIGGKNLPTRGTAEWLSFDVTDTVREWLLRRESNLGLEISIHCPCHTFQPNGDILENIHEVMEIKFKGVDNEDDHGRGDLGRLKKQKDHHNPHLILMMIPPHRLDNPGQGGQRKKRALDTNYCFRNLEENCCVRPLYIDFRQDLGWKWVHEPKGYYANFCSGPCPYLRSADTTHSTVLGLYNTLNPEASASPCCVPQDLEPLTILYYVGRTPKVEQLSNMVVKSCKCS DIP protein P10600 original sequence version: 125 DIP protein P10600 original sequence version: 171 3CAD3548D3AEA178 tgbr3_human Transforming growth factor beta receptor type 3 TGFBR3 Betaglycan Transforming growth factor beta receptor III protein protein human Homo sapiens Human MTSHYVIAIFALMSSCLATAGPEPGALCELSPVSASHPVQALMESFTVLSGCASRGTTGLPQEVHVLNLRTAGQGPGQLQREVTLHLNPISSVHIHHKSVVFLLNSPHPLVWHLKTERLATGVSRLFLVSEGSVVQFSSANFSLTAETEERNFPHGNEHLLNWARKEYGAVTSFTELKIARNIYIKVGEDQVFPPKCNIGKNFLSLNYLAEYLQPKAAEGCVMSSQPQNEEVHIIELITPNSNPYSAFQVDITIDIRPSQEDLEVVKNLILILKCKKSVNWVIKSFDVKGSLKIIAPNSIGFGKESERSMTMTKSIRDDIPSTQGNLVKWALDNGYSPITSYTMAPVANRFHLRLENNAEEMGDEEVHTIPPELRILLDPGALPALQNPPIRGGEGQNGGLPFPFPDISRRVWNEEGEDGLPRPKDPVIPSIQLFPGLREPEEVQGSVDIALSVKCDNEKMIVAVEKDSFQASGYSGMDVTLLDPTCKAKMNGTHFVLESPLNGCGTRPRWSALDGVVYYNSIVIQVPALGDSSGWPDGYEDLESGDNGFPGDMDEGDASLFTRPEIVVFNCSLQQVRNPSSFQEQPHGNITFNMELYNTDLFLVPSQGVFSVPENGHVYVEVSVTKAEQELGFAIQTCFISPYSNPDRMSHYTIIENICPKDESVKFYSPKRVHFPIPQADMDKKRFSFVFKPVFNTSLLFLQCELTLCTKMEKHPQKLPKCVPPDEACTSLDASIIWAMMQNKKTFTKPLAVIHHEAESKEKGPSMKEPNPISPPIFHGLDTLTVMGIAFAAFVIGALLTGALWYIYSHTGETAGRQQVPTSPPASENSSAAHSIGSTQSTPCSSSSTA AEC56C4477C003B2 tgfb3-tgfbr1 4 TGF-beta type I receptor 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 TGF-beta type II receptor 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:06:34 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg25Glu undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:06:35 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg94Glu undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:06:31 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Tyr90Ala Stoichiometry: 2.0 physical association physical association homomint mint f4a f4b t1 t2 sf1b tgfb3-tgfbr2-2 4 TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:06:20 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Tyr90Ala undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:06:17 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg94Glu undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:06:16 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg25Glu Stoichiometry: 2.1 TGF-beta type II receptor 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.2 direct interaction direct interaction homomint mint f4a f4b f4c t1 t2 sf1a tgfb3-tgfbr1-1 4 TGF-beta type I receptor 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified undefined mutation mutation disrupting interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:06:05 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Cys77Ser Stoichiometry: 2.0 TGF-beta type II receptor 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 physical association physical association homomint mint f4d t1 tgfb3-tgfbr3 4 TGF-beta type III receptor 8 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified direct interaction direct interaction homomint mint t2 f5 sf2 the endoglin-like domain of betaglycan, or BGe, was studied. tgfb3-1 3 TGF beta 3 7 weight by staining molecular weight estimation by staining weight by staining predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified Stoichiometry: 2.0 direct interaction direct interaction mint f2a f2b f2c The folding mixture, which contained the desired heterodimer, TGF-b3 WD, as well as wild-type and substituted homodimers, TGF-b3 WW and TGF-b3 DD, respectively, was then fractionated using high-resolution cation-exchange chromatography at pH 4.0 (Figure 2A). This separation yielded five major species, and as anticipated, three of these, b, d, and e, corresponded to reductantsensitive 25 kDa dimers (Figure 2B). The other two, a and c, corresponded to 12.5 kDa monomers (Figure 2B). The three dimers, as well as the two monomers were predicted to be positively charged under the experimental conditions, though reductions in the positive charge were expected for each arginine to glutamate substitution. Thus, peaks e, d, and b were predicted to correspond to the TGF-b3 WW, WD, and DD dimers, respectively, while peaks c and a, the TGF-b3 W and D monomers. tgfb3-tgfbr1-2 3 TGF-beta type II receptor 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified TGF-beta type I receptor 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain physical association physical association homomint mint To quantitate stoichiometries, the areas under the peaks were measured and compared with those for 2:2:1 and 1:1:1 TbRI:TbRII:TGF-b3 dimer complex calculated from the corresponding molar extinction coefficients at 280 nm. The results show that the relative integrated HPLC peak areas uncorrected for differences in extinction coefficients for the TbRI:TbRII:TGF-b3 WW complex, 0.099:0.45:1.00, closely match those expected for a 2:2:1 TbRI:TbRII:TGF-b complex, 0.085:0.41:1.00, whereas those for TGF-b3 WD complex, 0.043:0.16:1.00, match those expected for a 1:1:1 TbRI:TbRII: TGF-b complex, 0.043:0.20:1.00 (Table III). These results unambiguously demonstrate that the TGF-b3 WD heterodimer binds TbRII-ED and recruits TbRI-ED with an affinity indistinguishable from the TGF-b3 WT homodimer, but with one-half the stochiometry. 6d sf3 tiii P10600 increases activity of P36897 P10600 increases activity of P37173 tgfb3-tgfbr2-3 2 TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:47:33 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg94Glu undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:47:31 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg25Glu undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:47:28 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Tyr90Ala Stoichiometry: 2.1 TGF-beta type II receptor 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.2 direct interaction direct interaction f3a homomint mint The results showed that TGF-b3 WT, WW, and WD dimers formed detectable complexes with TbRII-ED, whereas TGF-b3 DD did not (Figure 3A). tgfb3-tgfbr1-3 2 TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:45:46 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Tyr90Ala undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:45:42 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg94Glu undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:45:41 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Arg25Glu Stoichiometry: 2.0 TGF-beta type I receptor 5 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 TGF-beta type II receptor 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 physical association physical association homomint mint The subsequent experiments focused on TbRI recruitment and were performed by titrating a fixed amount of TbRI extracellular domain, or TbRI-ED, with increasing amounts of TbRII-ED:TGF-b dimer complex. The TbRII:TGF-b complex was always added in a 2:1 molar ratio, regardless of whether the TbRII-ED was needed or not, to ensure that binding of TbRI-ED was not limited by TbRII-ED. The ligands that bound TbRII, TGF-b3 WT, WW, and WD, also bound and recruited TbRI-ED. TGF-b3 DD, which did not detectably bind TbRII, also failed to bind and recruit TbRI (Figure 3B). f3b f6c tgfb3-tgfbr1-4 1 TGF-beta type II receptor 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 TGF beta 3 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 TGF-beta type I receptor 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Stoichiometry: 1.0 physical association physical association homomint mint f6a f6b