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/ riou-2013-2 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation riou-2013-7 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation riou-2013-4 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation riou-2013-3 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 pull down pull down anti tag western anti tag western blot anti tag western Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation riou-2013-1 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip sequence tag sequence tag identification sequence tag Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation riou-2013-5 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation riou-2013-6 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation riou-2013-8 14-3-3 proteins interact with a hybrid prenyl-phosphorylation motif to inhibit G proteins. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Riou P., Kjær S., Garg R., Purkiss A., George R., Cain R.J., Bineva G., Reymond N., McColl B., Thompson A.J., O'Reilly N., McDonald N.Q., Parker P.J., Ridley A.J. Source of constructs obtained from the authors. Interactions identified using the non-specific 14-3-3 antibody were not captured. peter.parker@cancer.org.uk, anne.ridley@kcl.ac.uk Cell (0092-8674) 2013 Only protein-protein interactions imex curation 1433t_human 14-3-3 protein theta YWHAQ 14-3-3 protein tau 14-3-3 protein T-cell Protein HS1 protein protein human Homo sapiens Human MEKTELIQKAKLAEQAERYDDMATCMKAVTEQGAELSNEERNLLSVAYKNVVGGRRSAWRVISSIEQKTDTSDKKLQLIKDYREKVESELRSICTTVLELLDKYLIANATNPESKVFYLKMKGDYFRYLAEVACGDDRKQTIDNSQGAYQEAFDISKKEMQPTHPIRLGLALNFSVFYYEILNNPELACTLAKTAFDEAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDSAGEECDAAEGAEN rnd3_mouse Rho-related GTP-binding protein RhoE Rnd3 Arhe Rhoe Rho family GTPase 3 Rnd3 protein protein mouse Mus musculus Mouse MKERRASQKLSSKSIMDPNQNVKCKIVVVGDSQCGKTALLHVFAKDCFPENYVPTVFENYTASFEIDTQRIELSLWDTSGSPYYDNVRPLSYPDSDAVLICFDISRPETLDSVLKKWKGEIQEFCPNTKMLLVGCKSDLRTDVSTLVELSNHRQTPVSYDQGANMAKQIGAATYIECSALQSENSVRDIFHVATLACVNKTNKNVKRNKSQRATKRISHMPSRPELSAVATDLRKDKAKSCTVM 1433b_human 14-3-3 protein beta/alpha YWHAB Protein kinase C inhibitor protein 1 Protein 1054 protein protein human Homo sapiens Human MTMDKSELVQKAKLAEQAERYDDMAAAMKAVTEQGHELSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTERNEKKQQMGKEYREKIEAELQDICNDVLELLDKYLIPNATQPESKVFYLKMKGDYFRYLSEVASGDNKQTTVSNSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLNEESYKDSTLIMQLLRDNLTLWTSENQGDEGDAGEGEN 1433g_human 14-3-3 protein gamma YWHAG Protein kinase C inhibitor protein 1 protein protein human Homo sapiens Human MVDREQLVQKARLAEQAERYDDMAAAMKNVTELNEPLSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTSADGNEKKIEMVRAYREKIEKELEAVCQDVLSLLDNYLIKNCSETQYESKVFYLKMKGDYYRYLAEVATGEKRATVVESSEKAYSEAHEISKEHMQPTHPIRLGLALNYSVFYYEIQNAPEQACHLAKTAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDDDGGEGNN rhg05_human Rho GTPase-activating protein 5 ARHGAP5 RHOGAP5 Rho-type GTPase-activating protein 5 p190-B protein protein human Homo sapiens Human MMAKNKEPRPPSYTISIVGLSGTEKDKGNCGVGKSCLCNRFVRSKADEYYPEHTSVLSTIDFGGRVVNNDHFLYWGDIIQNSEDGVECKIHVIEQTEFIDDQTFLPHRSTNLQPYIKRAAASKLQSAEKLMYICTDQLGLEQDFEQKQMPEGKLNVDGFLLCIDVSQGCNRKFDDQLKFVNNLFVQLSKSKKPVIIAATKCDECVDHYLREVQAFASNKKNLLVVETSARFNVNIETCFTALVQMLDKTRSKPKIIPYLDAYKTQRQLVVTATDKFEKLVQTVRDYHATWKTVSNKLKNHPDYEEYINLEGTRKARNTFSKHIEQLKQEHIRKRREEYINTLPRAFNTLLPNLEEIEHLNWSEALKLMEKRADFQLCFVVLEKTPWDETDHIDKINDRRIPFDLLSTLEAEKVYQNHVQHLISEKRRVEMKEKFKKTLEKIQFISPGQPWEEVMCFVMEDEAYKYITEADSKEVYGRHQREIVEKAKEEFQEMLFEHSELFYDLDLNATPSSDKMSEIHTVLSEEPRYKALQKLAPDRESLLLKHIGFVYHPTKETCLSGQNCTDIKVEQLLASSLLQLDHGRLRLYHDSTNIDKVNLFILGKDGLAQELANEIRTQSTDDEYALDGKIYELDLRPVDAKSPYFLSQLWTAAFKPHGCFCVFNSIESLSFIGEFIGKIRTEASQIRKDKYMANLPFTLILANQRDSISKNLPILRHQGQQLANKLQCPFVDVPAGTYPRKFNETQIKQALRGVLESVKHNLDVVSPIPANKDLSEADLRIVMCAMCGDPFSVDLILSPFLDSHSCSAAQAGQNNSLMLDKIIGEKRRRIQITILSYHSSIGVRKDELVHGYILVYSAKRKASMGMLRAFLSEVQDTIPVQLVAVTDSQADFFENEAIKELMTEGEHIATEITAKFTALYSLSQYHRQTEVFTLFFSDVLEKKNMIENSYLSDNTRESTHQSEDVFLPSPRDCFPYNNYPDSDDDTEAPPPYSPIGDDVQLLPTPSDRSRYRLDLEGNEYPIHSTPNCHDHERNHKVPPPIKPKPVVPKTNVKKLDPNLLKTIEAGIGKNPRKQTSRVPLAHPEDMDPSDNYAEPIDTIFKQKGYSDEIYVVPDDSQNRIKIRNSFVNNTQGDEENGFSDRTSKSHGERRPSKYKYKSKTLFSKAKSYYRRTHSDASDDEAFTTSKTKRKGRHRGSEEDPLLSPVETWKGGIDNPAITSDQELDDKKMKKKTHKVKEDKKQKKKTKNFNPPTRRNWESNYFGMPLQDLVTAEKPIPLFVEKCVEFIEDTGLCTEGLYRVSGNKTDQDNIQKQFDQDHNINLVSMEVTVNAVAGALKAFFADLPDPLIPYSLHPELLEAAKIPDKTERLHALKEIVKKFHPVNYDVFRYVITHLNRVSQQHKINLMTADNLSICFWPTLMRPDFENREFLSTTKIHQSVVETFIQQCQFFFYNGEIVETTNIVAPPPPSNPGQLVEPMVPLQLPPPLQPQLIQPQLQTDPLGII GTPase-activating protein for Rho family members. May play a role in the reduction of the p21rasGTPase-activating potential of p120GAP mint homomint 1433e_human 14-3-3 protein epsilon YWHAE protein protein human Homo sapiens Human MDDREDLVYQAKLAEQAERYDEMVESMKKVAGMDVELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEENKGGEDKLKMIREYRQMVETELKLICCDILDVLDKHLIPAANTGESKVFYYKMKGDYHRYLAEFATGNDRKEAAENSLVAYKAASDIAMTELPPTHPIRLGLALNFSVFYYEILNSPDRACRLAKAAFDDAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDMQGDGEEQNKEALQDVEDENQ 1433z_human 14-3-3 protein zeta/delta YWHAZ Protein kinase C inhibitor protein 1 protein protein human Homo sapiens Human MDKNELVQKAKLAEQAERYDDMAACMKSVTEQGAELSNEERNLLSVAYKNVVGARRSSWRVVSSIEQKTEGAEKKQQMAREYREKIETELRDICNDVLSLLEKFLIPNASQAESKVFYLKMKGDYYRYLAEVAAGDDKKGIVDQSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDTQGDEAEAGEGGEN 1433f_human 14-3-3 protein eta YWHAH YWHA1 Protein AS1 protein protein human Homo sapiens Human MGDREQLLQRARLAEQAERYDDMASAMKAVTELNEPLSNEDRNLLSVAYKNVVGARRSSWRVISSIEQKTMADGNEKKLEKVKAYREKIEKELETVCNDVLSLLDKFLIKNCNDFQYESKVFYLKMKGDYYRYLAEVASGEKKNSVVEASEAAYKEAFEISKEQMQPTHPIRLGLALNFSVFYYEIQNAPEQACLLAKQAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDEEAGEGN 1433s_human 14-3-3 protein sigma Stratifin Epithelial cell marker protein 1 HME1 SFN protein protein human Homo sapiens Human MERASLIQKAKLAEQAERYEDMAAFMKGAVEKGEELSCEERNLLSVAYKNVVGGQRAAWRVLSSIEQKSNEEGSEEKGPEVREYREKVETELQGVCDTVLGLLDSHLIKEAGDAESRVFYLKMKGDYYRYLAEVATGDDKKRIIDSARSAYQEAMDISKKEMPPTNPIRLGLALNFSVFHYEIANSPEEAISLAKTTFDEAMADLHTLSEDSYKDSTLIMQLLRDNLTLWTADNAGEEGGEAPQEPQS rnd3_human Rho-related GTP-binding protein RhoE RND3 ARHE RHO8 RHOE Rho family GTPase 3 Rnd3 Rho-related GTP-binding protein Rho8 Protein MemB protein protein human Homo sapiens Human MKERRASQKLSSKSIMDPNQNVKCKIVVVGDSQCGKTALLHVFAKDCFPENYVPTVFENYTASFEIDTQRIELSLWDTSGSPYYDNVRPLSYPDSDAVLICFDISRPETLDSVLKKWKGEIQEFCPNTKMLLVGCKSDLRTDVSTLVELSNHRQTPVSYDQGANMAKQIGAATYIECSALQSENSVRDIFHVATLACVNKTNKNVKRNKSQRATKRISHMPSRPELSAVATDLRKDKAKSCTVM rap1a_human Ras-related protein Rap-1A GTP-binding protein smg p21A Ras-related protein Krev-1 C21KG G-22K RAP1A KREV1 protein protein human Homo sapiens Human MREYKLVVLGSGGVGKSALTVQFVQGIFVEKYDPTIEDSYRKQVEVDCQQCMLEILDTAGTEQFTAMRDLYMKNGQGFALVYSITAQSTFNDLQDLREQILRVKDTEDVPMILVGNKCDLEDERVVGKEQGQNLARQWCNCAFLESSAKSKINVNEIFYDLVRQINRKTPVEKKKPKKKSCLLL 1433g_chlae_protein 14-3-3 protein gamma protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey An amino acid sequence which is not represented in UniProt Similar to P61982 1433b_chlae_protein 14-3-3 protein beta/alpha protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey An amino acid sequence which is not represented in UniProt 1433s_chlae_protein 14-3-3 protein sigma protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey Similar to P31947 An amino acid sequence which is not represented in UniProt 1433e_chlae_protein protein protein chlae Chlorocebus aethiops Cercopithecus aethiops Grivet Green monkey Similar to P62260 rnd3-ywhaq 1434296 n/a 1434304 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role bait bait region 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 false physical association physical association false false false Supp. Fig. 1A, 1B staurosporine calyculin A rnd3-ywhab 1434296 n/a 1434305 unspecified role unspecified role bait bait region 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 false n/a 1434306 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 1A, Supp. Fig 1A, 1B Staurosporine calyculin A Calf intestinal phosphatase rnd3-ywhag 1434296 n/a 1434305 unspecified role unspecified role bait bait region 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 false n/a 1434307 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. Fig. 1A, 1B calyculin A Staurosporine ywhag-rnd3 1434296 n/a 1434307 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role prey prey region 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 false physical association physical association false false false Supp. Fig. 1A ywhaq-rnd3 1434296 n/a 1434304 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role prey prey region 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 false physical association physical association false false false Supp. Fig. 1A ywhab-rnd3 1434296 n/a 1434305 unspecified role unspecified role prey prey region 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 false n/a 1434306 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. Fig 1A rnd3-arhgap5 1434296 n/a 1434308 unspecified role unspecified role prey prey p region required to bind necessary binding region required to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role bait bait region 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 false P61588:p.Thr55Val mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Supp. Fig. 3B arhgap5-rnd3 1434296 n/a 1434308 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false p region required to bind necessary binding region required to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role prey prey region 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 false physical association physical association false false false Supp. Fig. 3C Interaction not observed in presence of 14-3-3beta protein (YWHAB) rnd3-ywhab-1 1434296 n/a 1434306 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role bait bait region 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 false P61588:p.[Ser210Ala;Thr214Ala;Ser218Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ser-210 (possible) opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Supp. Fig. 3C ywhae-rnd3 1434296 n/a 1434305 unspecified role unspecified role prey prey region 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 false n/a 1434309 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. Fig. 1A rnd3-ywhae 1434296 n/a 1434305 unspecified role unspecified role bait bait region 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 false n/a 1434309 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 1A Supp. Fig. 1A, 1B staurosporine calyculin A rnd3-ywhaz 1434296 n/a 1434305 unspecified role unspecified role bait bait region 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 false n/a 1434310 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 1A Supp. Fig. 1A, 1B staurosporine calyculin A ywhah-rnd3 1434296 n/a 1434305 unspecified role unspecified role prey prey region 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 false n/a 1434311 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. Fig. 1A rnd3-ywhah 1434296 n/a 1434311 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role bait bait region 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 false physical association physical association false false false Supp. Fig. 1A, 1B staurosporine calyculin A ywhaz-rnd3 1434296 n/a 1434310 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role prey prey region 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 false physical association physical association false false false Supp. Fig. 1A rnd3-ywhaz 1434296 n/a 1434310 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1434305 unspecified role unspecified role bait bait region 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 false physical association physical association false false false Supp. Fig. 1A, 1B calyculin A Staurosporine sfn-rnd3-1 1434296 n/a 1434305 unspecified role unspecified role prey prey region 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 false n/a 1434312 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Supp. Fig. 1A rnd3-ywhaz-1 1434297 n/a 1434313 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false cys-241 (required) farnres farnesylated residue FarnRes Farnesyl Farnesylation certain certain sequence position certain certain certain sequence position certain false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 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 false region 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) 1434417 1434414 direct interaction direct interaction false false false 1434297 Table 2 rnd3-ywhaz-2 1434297 n/a 1434313 unspecified role unspecified role bait bait cys-241 (required) ipcys isoprenylated cysteine IpCys certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) 1434425 1434423 direct interaction direct interaction false false false 1434297 Table 2 rnd3-ywhaz-3 1434297 n/a 1434313 unspecified role unspecified role bait bait ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false cys-241 (required) sgergercys S-geranylgeranyl-L-cysteine 2-amino-3-(3,7,11,15-tetramethyl-2,6,10,14-hexadecatetraenylthio)propanoic acid LIPID S-geranylgeranyl cysteine S-geranylgeranyl-L-cysteine (2R)-2-amino-3-([(2E,6E,10Z)-12-hydroxy-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]sulfanyl)propanoic acid Geranyl-geranyl GeranylGeranyl S-geranylgeranyl SGergerCys geranylgeranylc certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) 1434434 1434431 direct interaction direct interaction false false false 1434297 Table 2 rnd3-ywhaz-4 1434297 n/a 1434313 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false cys-241 geranylated (required) modres protein modification ModRes certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) 1434443 1434441 direct interaction direct interaction false false false 1434297 Table 2 rnd3-ywhaz-5 1434297 n/a 1434313 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1434297 Table 2 rap1a-ywhaz 1434297 n/a 1434314 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false cys-181 sgergercys S-geranylgeranyl-L-cysteine 2-amino-3-(3,7,11,15-tetramethyl-2,6,10,14-hexadecatetraenylthio)propanoic acid LIPID S-geranylgeranyl cysteine S-geranylgeranyl-L-cysteine (2R)-2-amino-3-([(2E,6E,10Z)-12-hydroxy-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]sulfanyl)propanoic acid Geranyl-geranyl GeranylGeranyl S-geranylgeranyl SGergerCys geranylgeranylc certain certain sequence position certain certain certain sequence position certain false n/a 1434310 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) 1434459 1434456 direct interaction direct interaction false false false 1434297 Fig. 7C Removal of the geranylgeranylated modification resulted in the Rap1A peptide having micromolar affinity for the 14-3-3 protein. rnd3-ywhab-3 1434298 n/a 1434313 unspecified role unspecified role bait bait ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false cys-241 (required) farnres farnesylated residue FarnRes Farnesyl Farnesylation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434306 unspecified role unspecified role prey prey 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) direct interaction direct interaction false false false Fig. 5B rnd3-ywhaz-6 1434298 n/a 1434313 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false cys-241 (required) farnres farnesylated residue FarnRes Farnesyl Farnesylation certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 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 false region 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) 1434477 1434472 direct interaction direct interaction false false false Fig. 7A rnd3-ywhaz-7 1434298 n/a 1434313 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false cys-241 (required) ipcys isoprenylated cysteine IpCys certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) 1434485 1434481 direct interaction direct interaction false false false Fig. 7A rnd3-ywhaz-8 1434298 n/a 1434313 unspecified role unspecified role bait bait ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false cys-241 (required) sgergercys S-geranylgeranyl-L-cysteine 2-amino-3-(3,7,11,15-tetramethyl-2,6,10,14-hexadecatetraenylthio)propanoic acid LIPID S-geranylgeranyl cysteine S-geranylgeranyl-L-cysteine (2R)-2-amino-3-([(2E,6E,10Z)-12-hydroxy-3,7,11-trimethyldodeca-2,6,10-trien-1-yl]sulfanyl)propanoic acid Geranyl-geranyl GeranylGeranyl S-geranylgeranyl SGergerCys geranylgeranylc certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) 1434494 1434492 direct interaction direct interaction false false false Fig. 7A rnd3-ywhaz-9 1434298 n/a 1434313 unspecified role unspecified role bait bait cys-241 geranylated (required) modres protein modification ModRes certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 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 false region 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) 1434504 1434500 direct interaction direct interaction false false false Fig. 7A ywhab-rnd3-1 1434299 n/a 1434306 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 1434305 unspecified role unspecified role prey prey region 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 false physical association physical association false false false Fig. 5A ywhab-rnd3-2 1434299 n/a 1434306 competitor competitor competitor prey prey ser-240 (required) oglycoser O-glycosyl-L-serine OGlycoSer certain certain sequence position certain certain certain sequence position certain false cys-241 (required) farnres farnesylated residue FarnRes Farnesyl Farnesylation certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434306 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 1434305 unspecified role unspecified role prey prey region 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 false association association false false false Fig. 5A rnd3-1433b_chlae_p-1 1434300 n/a 1434315 unspecified role unspecified role prey prey n/a 1434316 unspecified role unspecified role prey prey n/a 1434305 unspecified role unspecified role bait bait region 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 false n/a 1434317 unspecified role unspecified role prey prey n/a 1434318 unspecified role unspecified role prey prey association association false false false Supp. Table 1 rnd3-ywhab-7 1434301 n/a 1434313 unspecified role unspecified role neutral component neutral component ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false cys-241 (required) farnres farnesylated residue FarnRes Farnesyl Farnesylation certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434310 unspecified role unspecified role neutral component neutral component 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 false region 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) 1434534 1434530 direct interaction direct interaction false false false 1434301 Fig. 5C Table 1, Supp. Fig. 5 Defarnesylation of Rnd3 increased the Kd to 1 uM. rnd3-ywhab-8 1434302 n/a 1434313 unspecified role unspecified role neutral component neutral component cys-241 (required) farnres farnesylated residue FarnRes Farnesyl Farnesylation certain certain sequence position certain certain certain sequence position certain false ser-240 (required) nphosres N-phosphorylated residue NPhosRes certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 2.0 n/a 1434310 unspecified role unspecified role neutral component neutral component 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 false region 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) Stoichiometry: 2.0 direct interaction direct interaction false false false 1434302 Supp. Table 4 2.3A Rworking = 0.24 Rfree = 0.27 P63104 decreases activity of P61587 ywhab-rnd3-3 1434303 n/a 1434313 unspecified role unspecified role prey prey P61587:p.[Ser7Ala;Ser11Ala;Ser210Ala;Thr214Ala;Ser218Ala;Ser222Ala;Ser240Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region 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 false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1434306 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supp. Fig. 1C