mbinfo MBInfo http://www.mechanobio.info/ Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. J. Biol. Chem. (0021-9258) 2008 imex curation Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp 15-Nov-2012: Contacted by pporras. Only protein-protein interactions human-platelet Homo sapiens platelet, thrombocytes platelet Platelet, thrombocyte anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp Accepted 2012-NOV-02 by ORCHARD 15-Nov-2012: Contacted by pporras. J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. J. Biol. Chem. (0021-9258) 2008 imex curation Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp 15-Nov-2012: Contacted by pporras. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp Accepted 2012-NOV-02 by ORCHARD 15-Nov-2012: Contacted by pporras. J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. J. Biol. Chem. (0021-9258) 2008 imex curation Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp 15-Nov-2012: Contacted by pporras. Only protein-protein interactions in vitro In vitro pull down pull down ms participant Identification by mass spectrometry ms participant Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp Accepted 2012-NOV-02 by ORCHARD 15-Nov-2012: Contacted by pporras. J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets. J. Biol. Chem. (0021-9258) 2008 imex curation Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp 15-Nov-2012: Contacted by pporras. 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 Higashi T., Ikeda T., Shirakawa R., Kondo H., Kawato M., Horiguchi M., Okuda T., Okawa K., Fukai S., Nureki O., Kita T., Horiuchi H. The binding between mDia1 and RhoA (described in the text) could not be curated since it could not be supported by a figure reference. Interaction detection : GST pull down. The source of RhoA specified by the author and the source of DAAM1 and mDIa1 (both endogenous) inferred from the cells (human platelets) used. horiuchi@kuhp.kyoto-u.ac.jp Accepted 2012-NOV-02 by ORCHARD 15-Nov-2012: Contacted by pporras. J. Biol. Chem. (0021-9258) 2008 Only protein-protein interactions imex curation daam1_human Disheveled-associated activator of morphogenesis 1 DAAM1 KIAA0666 protein protein human Homo sapiens Human MAPRKRGGRGISFIFCCFRNNDHPEITYRLRNDSNFALQTMEPALPMPPVEELDVMFSELVDELDLTDKHREAMFALPAEKKWQIYCSKKKDQEENKGATSWPEFYIDQLNSMAARKSLLALEKEEEEERSKTIESLKTALRTKPMRFVTRFIDLDGLSCILNFLKTMDYETSESRIHTSLIGCIKALMNNSQGRAHVLAHSESINVIAQSLSTENIKTKVAVLEILGAVCLVPGGHKKVLQAMLHYQKYASERTRFQTLINDLDKSTGRYRDEVSLKTAIMSFINAVLSQGAGVESLDFRLHLRYEFLMLGIQPVIDKLREHENSTLDRHLDFFEMLRNEDELEFAKRFELVHIDTKSATQMFELTRKRLTHSEAYPHFMSILHHCLQMPYKRSGNTVQYWLLLDRIIQQIVIQNDKGQDPDSTPLENFNIKNVVRMLVNENEVKQWKEQAEKMRKEHNELQQKLEKKERECDAKTQEKEEMMQTLNKMKEKLEKETTEHKQVKQQVADLTAQLHELSRRAVCASIPGGPSPGAPGGPFPSSVPGSLLPPPPPPPLPGGMLPPPPPPLPPGGPPPPPGPPPLGAIMPPPGAPMGLALKKKSIPQPTNALKSFNWSKLPENKLEGTVWTEIDDTKVFKILDLEDLERTFSAYQRQQDFFVNSNSKQKEADAIDDTLSSKLKVKELSVIDGRRAQNCNILLSRLKLSNDEIKRAILTMDEQEDLPKDMLEQLLKFVPEKSDIDLLEEHKHELDRMAKADRFLFEMSRINHYQQRLQSLYFKKKFAERVAEVKPKVEAIRSGSEEVFRSGALKQLLEVVLAFGNYMNKGQRGNAYGFKISSLNKIADTKSSIDKNITLLHYLITIVENKYPSVLNLNEELRDIPQAAKVNMTELDKEISTLRSGLKAVETELEYQKSQPPQPGDKFVSVVSQFITVASFSFSDVEDLLAEAKDLFTKAVKHFGEEAGKIQPDEFFGIFDQFLQAVSEAKQENENMRKKKEEEERRARMEAQLKEQRERERKMRKAKENSEESGEFDDLVSALRSGEVFDKDLSKLKRNRKRITNQMTDSSRERPITKLNF E82D9892390254B6 rhob_human Rho-related GTP-binding protein RhoB Rho cDNA clone 6 RHOB ARH6 ARHB protein protein human Homo sapiens Human MAAIRKKLVVVGDGACGKTCLLIVFSKDEFPEVYVPTVFENYVADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSVDSPDSLENIPEKWVPEVKHFCPNVPIILVANKKDLRSDEHVRTELARMKQEPVRTDDGRAMAVRIQAYDYLECSAKTKEGVREVFETATRAALQKRYGSQNGCINCCKVL CCE6FD53AE00CD83 rhoc_human Rho-related GTP-binding protein RhoC Rho cDNA clone 9 RHOC ARH9 ARHC protein protein human Homo sapiens Human MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYIADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKKDLRQDEHTRRELAKMKQEPVRSEEGRDMANRISAFGYLECSAKTKEGVREVFEMATRAGLQVRKNKRRRGCPIL A193DA581560F131 diap1_human Protein diaphanous homolog 1 DIAPH1 DIAP1 Diaphanous-related formin-1 protein protein human Homo sapiens Human MEPPGGSLGPGRGTRDKKKGRSPDELPSAGGDGGKSKKFTLKRLMADELERFTSMRIKKEKEKPNSAHRNSSASYGDDPTAQSLQDVSDEQVLVLFEQMLLDMNLNEEKQQPLREKDIIIKREMVSQYLYTSKAGMSQKESSKSAMMYIQELRSGLRDMPLLSCLESLRVSLNNNPVSWVQTFGAEGLASLLDILKRLHDEKEETAGSYDSRNKHEIIRCLKAFMNNKFGIKTMLETEEGILLLVRAMDPAVPNMMIDAAKLLSALCILPQPEDMNERVLEAMTERAEMDEVERFQPLLDGLKSGTTIALKVGCLQLINALITPAEELDFRVHIRSELMRLGLHQVLQDLREIENEDMRVQLNVFDEQGEEDSYDLKGRLDDIRMEMDDFNEVFQILLNTVKDSKAEPHFLSILQHLLLVRNDYEARPQYYKLIEECISQIVLHKNGADPDFKCRHLQIEIEGLIDQMIDKTKVEKSEAKAAELEKKLDSELTARHELQVEMKKMESDFEQKLQDLQGEKDALHSEKQQIATEKQDLEAEVSQLTGEVAKLTKELEDAKKEMASLSAAAITVPPSVPSRAPVPPAPPLPGDSGTIIPPPPAPGDSTTPPPPPPPPPPPPPLPGGVCISSPPSLPGGTAISPPPPLSGDATIPPPPPLPEGVGIPSPSSLPGGTAIPPPPPLPGSARIPPPPPPLPGSAGIPPPPPPLPGEAGMPPPPPPLPGGPGIPPPPPFPGGPGIPPPPPGMGMPPPPPFGFGVPAAPVLPFGLTPKKLYKPEVQLRRPNWSKLVAEDLSQDCFWTKVKEDRFENNELFAKLTLTFSAQTKTSKAKKDQEGGEEKKSVQKKKVKELKVLDSKTAQNLSIFLGSFRMPYQEIKNVILEVNEAVLTESMIQNLIKQMPEPEQLKMLSELKDEYDDLAESEQFGVVMGTVPRLRPRLNAILFKLQFSEQVENIKPEIVSVTAACEELRKSESFSNLLEITLLVGNYMNAGSRNAGAFGFNISFLCKLRDTKSTDQKMTLLHFLAELCENDYPDVLKFPDELAHVEKASRVSAENLQKNLDQMKKQISDVERDVQNFPAATDEKDKFVEKMTSFVKDAQEQYNKLRMMHSNMETLYKELGEYFLFDPKKLSVEEFFMDLHNFRNMFLQAVKENQKRRETEEKMRRAKLAKEKAEKERLEKQQKREQLIDMNAEGDETGVMDSLLEALQSGAAFRRKRGPRQANRKAGCAVTSLLASELTKDDAMAAVPAKVSKNSETFPTILEEAKELVGRAS 9C8273DE4748564F rhof_human Rho-related GTP-binding protein RhoF RHOF ARHF RIF Rho family GTPase Rif Rho in filopodia protein protein human Homo sapiens Human MDAPGALAQTAAPGPGRKELKIVIVGDGGCGKTSLLMVYSQGSFPEHYAPSVFEKYTASVTVGSKEVTLNLYDTAGQEDYDRLRPLSYQNTHLVLICYDVMNPTSYDNVLIKWFPEVTHFCRGIPMVLIGCKTDLRKDKEQLRKLRAAQLEPITYMQGLSACEQIRAALYLECSAKFRENVEDVFREAAKVALSALKKAQRQKKRRLCLLL AF82AE0CEDC34FAD rhod_human Rho-related GTP-binding protein RhoD RHOD ARHD Rho-related protein HP1 protein protein human Homo sapiens Human MTAAQAAGEEAPPGVRSVKVVLVGDGGCGKTSLLMVFADGAFPESYTPTVFERYMVNLQVKGKPVHLHIWDTAGQDDYDRLRPLFYPDASVLLLCFDVTSPNSFDNIFNRWYPEVNHFCKKVPIIVVGCKTDLCKDKSLVNKLRRNGLEPVTYHRGQEMARSVGAVAYLECSARLHDNVHAVFQEAAEVALSSRGRNFWRRITQGFCVVT 7B8A7AE8933C78B0 rhoa_human Transforming protein RhoA Rho cDNA clone 12 ARHA RHO12 RHOA ARH12 protein protein human Homo sapiens Human MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVADIEVDGKQVELALWDTAGQEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVKHFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKTKDGVREVFEMATRAALQARRGKKKSGCLVL DIP protein P61586 original sequence version: 86 DIP protein P61586 original sequence version: 123 DIP protein P61586 original sequence version: 57 DIP protein P61586 original sequence version: 88 DIP protein P61586 original sequence version: 87 DIP protein P61586 original sequence version: 85 DIP protein P61586 original sequence version: 125 DIP protein P61586 original sequence version: 132 C4DA2DC31FF858BC rhoa-daam1-1 3 11 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey human-platelet Homo sapiens platelet, thrombocytes platelet Platelet, thrombocyte 5 unspecified role unspecified role prey prey human-platelet Homo sapiens platelet, thrombocytes platelet Platelet, thrombocyte association association Fig 1A. DAAM1 and mDia1 associated only with GTP-loaded RhoA and not with GDP-loaded RhoA. daam1-rhoa 4 11 unspecified role unspecified role unspecified role unspecified role region 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role unspecified role unspecified role n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 1E. DAAM1 bound to GTP-loaded RhoA with a Kd value of 4nM. DAAM1 showed a weaker association with GDP-loaded RhoA (data not shown). daam1-daam1-1 2 5 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 c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9Y4D1:p.Phe1046Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F A Wed May 02 20:03:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe1046ala' Tue Jun 30 18:15:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe1046Ala' ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9Y4D1:p.Ala229Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain A D Wed May 02 17:40:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala229asp' Tue Jun 30 16:52:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala229Asp' region 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 ecoli Escherichia coli (strain K12) 26 29 direct interaction direct interaction Fig 4A, 5A. diaph1-diaph1-9 2 8 unspecified role unspecified role bait bait c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 O60610:p.Leu1203Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L A Wed May 02 16:27:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu1203ala' Tue Jun 30 17:57:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu1203Ala' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey O60610:p.Met256Asp mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain M D Wed May 02 17:05:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'met256asp' Tue Jun 30 16:26:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Met256Asp' n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) 34 39 direct interaction direct interaction Fig 4B, 5B. rhoa-daam1-2 2 11 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 6A (upper panel). DAAM1 interacted only to GTPgammaS-loaded RhoA, not to GDP-loaded RhoA. rhob-daam1 2 5 unspecified role unspecified role prey prey region 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 n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 6A (upper panel). DAAM1 interacted only with the GTPgammaS-loaded RhoB, not with GDP-loaded RhoB. rhoc-daam1 2 7 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 ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role prey prey region 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 n-terminal region 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 Fig 6A (upper panel). DAAM1 interacted only with the GTPgammaS-loaded RhoC, not with the GDP-loaded RhoC. rhod-daam1 2 5 unspecified role unspecified role prey prey n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 6A (upper panel). DAAM1 interacted only with the GTPgammaS-loaded RhoD, not with the GDP-loaded RhoD. rhoa-diaph1-2 2 8 unspecified role unspecified role prey prey n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) 11 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 6A (lower panel). mDia1 interacted only with the GTPgammaS-loaded RhoA, not with the GDP-loaded RhoA. rhob-diaph1 2 6 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region 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 n-terminal region 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 Fig 6A (lower panel). mDia1 interacted only with the GTPgammaS-loaded RhoB, not with the GDP-loaded RhoB. rhoc-diaph1 2 8 unspecified role unspecified role prey prey n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) 7 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 6A (lower panel). mDia1 interacted only with the GTPgammaS-loaded RhoC, not with the GDP-loaded RhoC. rhod-diaph1 2 8 unspecified role unspecified role prey prey n-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig 6A (lower panel). mDia1 interacted only with the GTPgammaS-loaded RhoD, not with the GDP-loaded RhoD. rhof-diaph1 2 9 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region 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 region 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 Fig 6A (lower panel). mDia1 interacted only with the GTPgammaS-loaded RhoF, not with the GDP-loaded RhoF. daam1-rhoa-2 1 11 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association Fig 7A. RhoA coprecipitated with DAAM1 strongly in thrombin-stimulated platelets and weakly in unstimulated cells. Thrombin diaph1-rhoa-3 4 11 unspecified role unspecified role unspecified role unspecified role region 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role unspecified role unspecified role n-terminal region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain region 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Referenced in text. mDia1 had a higher affinity for GTP-bound RhoA with a Kd of 4.8nM, but had a lower affinity for GDP-bound RhoA with a Kd of 430nM.