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 Bivalent binding drives the formation of the Grb2-Gab1 signaling complex in a noncooperative manner. Bivalent binding drives the formation of the Grb2-Gab1 signaling complex in a noncooperative manner. FEBS J. (1742-464X) 2012 McDonald CB., Bhat V., Mikles DC., Deegan BJ., Seldeen KL., Farooq A. in vitro In vitro light scattering light scattering predetermined predetermined participant predetermined McDonald CB., Bhat V., Mikles DC., Deegan BJ., Seldeen KL., Farooq A. FEBS J. (1742-464X) 2012 Bivalent binding drives the formation of the Grb2-Gab1 signaling complex in a noncooperative manner. Bivalent binding drives the formation of the Grb2-Gab1 signaling complex in a noncooperative manner. FEBS J. (1742-464X) 2012 McDonald CB., Bhat V., Mikles DC., Deegan BJ., Seldeen KL., Farooq A. in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined McDonald CB., Bhat V., Mikles DC., Deegan BJ., Seldeen KL., Farooq A. FEBS J. (1742-464X) 2012 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 83A4B0BA1B248DC4 gab1_human GRB2-associated-binding protein 1 Growth factor receptor bound protein 2-associated protein 1 GRB2-associated binder 1 GAB1 protein protein human Homo sapiens Human MSGGEVVCSGWLRKSPPEKKLKRYAWKRRWFVLRSGRLTGDPDVLEYYKNDHAKKPIRIIDLNLCQQVDAGLTFNKKEFENSYIFDINTIDRIFYLVADSEEEMNKWVRCICDICGFNPTEEDPVKPPGSSLQAPADLPLAINTAPPSTQADSSSATLPPPYQLINVPPHLETLGIQEDPQDYLLLINCQSKKPEPTRTHADSAKSTSSETDCNDNVPSHKNPASSQSKHGMNGFFQQQMIYDSPPSRAPSASVDSSLYNLPRSYSHDVLPKVSPSSTEADGELYVFNTPSGTSSVETQMRHVSISYDIPPTPGNTYQIPRTFPEGTLGQTSKLDTIPDIPPPRPPKPHPAHDRSPVETCSIPRTASDTDSSYCIPTAGMSPSRSNTISTVDLNKLRKDASSQDCYDIPRAFPSDRSSSLEGFHNHFKVKNVLTVGSVSSEELDENYVPMNPNSPPRQHSSSFTEPIQEANYVPMTPGTFDFSSFGMQVPPPAHMGFRSSPKTPPRRPVPVADCEPPPVDRNLKPDRKVKPAPLEIKPLPEWEELQAPVRSPITRSFARDSSRFPMSPRPDSVHSTTSSSDSHDSEENYVPMNPNLSSEDPNLFGSNSLDGGSSPMIKPKGDKQVEYLDLDLDSGKSTPPRKQKSSGSGSSVADERVDYVVVDQQKTLALKSTREAWTDGRQSTESETPAKSVK E0FAC78772B6677B gab1-grb2-4 2 GRB2 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 2.1 GAB1 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 f4 t3 Our analysis reveals that Grb2 binds to the wildtype PR domain (PR_WT), containing both G1 and G2 sites, with a 2:1 stoichiometry (Table 3). Coupled with previous data [18, 28, 29], the most straightforward interpretation of this finding is that Grb2 binds in a bivalent manner to Gab1 to form the Grb2-Gab1 complex with a 2:1 stoichiometry and that this interaction is mediated through the binding of cSH3 domain within each Grb2 molecule to each of the two distinct RXXK motifs (G1 and G2) within Gab1 gab1-grb2-5 2 GAB1-mG2 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Q13480:p.Arg521Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R A Tue Jul 12 10:18:22 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 18:05:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg521ala' Tue Jun 30 16:36:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg521Ala' his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q13480:p.Lys524Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Tue Jul 12 10:16:38 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 15:42:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys524ala' Tue Jun 30 18:55:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys524Ala' Q13480:p.Asn522Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N A Tue Jul 12 10:16:37 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 19:20:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn522ala' Tue Jun 30 17:22:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn522Ala' binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Q13480:p.Leu523Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain L A Tue Jul 12 10:16:43 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 16:54:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu523ala' Tue Jun 30 16:18:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu523Ala' GRB2 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction homomint mint f4 t3 our data indicate that binding of Grb2 to the PR_WT domain occurs with an affinity that is greater than that observed for its binding to G1 site but lower than G2 site, or essentially an average of the two constituent affinity terms (Table 3). gab1-grb2-6 2 GAB1-mG1 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Q13480:p.Pro346Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain P A Tue Jul 12 10:16:45 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 16:10:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro346ala' Tue Jun 30 16:45:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro346Ala' Q13480:p.Lys347Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain K A Tue Jul 12 10:15:32 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 17:06:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys347ala' Tue Jun 30 18:35:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys347Ala' Q13480:p.Arg344Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain R A Tue Jul 12 10:15:31 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 17:46:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg344ala' Tue Jun 30 19:51:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg344Ala' binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Q13480:p.Pro345Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain P A Tue Jul 12 10:16:09 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 15:30:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro345ala' Tue Jun 30 17:50:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro345Ala' his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined GRB2 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction homomint mint f4 t3 our data indicate that binding of Grb2 to the PR_WT domain occurs with an affinity that is greater than that observed for its binding to G1 site but lower than G2 site, or essentially an average of the two constituent affinity terms (Table 3). grb2-4 1 GRB2 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 2.0 direct interaction direct interaction f2 t1 mint we generated Grb2_F61A, Grb2_F182A and Grb2_R207A single-alanine mutant constructs and analyzed and compared their hydrodynamic properties to the wildtype Grb2_WT construct (Table 1). To our surprise, all three mutant constructs of Grb2 failed to shift the dimer-monomer equilibrium in favor of the latter