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/ becker-2004a-3 Preprotein recognition by the Toc complex. in vitro In vitro biochemical biochemical western blot western blot Immuno blot Immuno blot Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. It is not clear from the text how the recombinant Toc proteins were produced. The curator has assumed the proetins were expressed in ecoli and subsequently purified and used in the assay. The details of the method used to produce Toc protein containing liposomes was obtained from the reference provided Schleiff et al (2003) (PMID: 12665619) in the text. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation becker-2004a-1 Preprotein recognition by the Toc complex. arath-leaf Arabidopsis thaliana leaf leaf leaf anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation becker-2004a-5 Preprotein recognition by the Toc complex. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. It is not clear from the text how the recombinant Toc159 protein was produced. The curator has assumed the proetins were expressed in ecoli and subsequently purified and used in the assay. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation becker-2004a-2 Preprotein recognition by the Toc complex. arath-leaf Arabidopsis thaliana leaf leaf leaf far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation becker-2004a-4 Preprotein recognition by the Toc complex. in vitro In vitro pull down pull down immunostaining immunostaining Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. It is not clear from the text how the recombinant Toc159 protein was produced. The curator has assumed the proetins were expressed in ecoli and subsequently purified and used in the assay. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation becker-2004a-6 Preprotein recognition by the Toc complex. in vitro In vitro gtpase assay gtpase assay GTPase gtp hydrolisis gtp hydrolisis GTPase autoradiography autoradiography Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. It is not clear from the text how the recombinant Toc159 protein was produced. The curator has assumed the proetins were expressed in ecoli and subsequently purified and used in the assay. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation becker-2004a-7 Preprotein recognition by the Toc complex. in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati weight by staining molecular weight estimation by staining weight by staining Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation becker-2004a-8 Preprotein recognition by the Toc complex. in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati western blot western blot Immuno blot Immuno blot Becker T., Jelic M., Vojta A., Radunz A., Soll J., Schleiff E. Radio-labelled pSSU protein was detected by autoradiography whereas Toc159 or Toc34 proteins were detected by western blotting. schleiff@botanik.biologie.uni-muenchen.de EMBO J. (0261-4189) 2004 Only protein-protein interactions imex curation toc34_arath Translocase of chloroplast 34, chloroplastic TOC34 OEP34 MUG13.14 At5g05000 PPI3 Plastid protein import 3 34 kDa chloroplast outer envelope protein GTP-binding protein OEP34 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAALQTLREWIGIQQFPPATQSKLLEILGKYKEEDVSSLTVLVMGKGGVGKSSTVNSVIGEKAAAVSTFQSEGLRPTLVSRTRSGFTLNIIDTPGLIEGGYVNDQAINIIKRFLLNMTIDVLLYVDRLDVYRVDDLDRQVVGAITDAFGKEIWKKSALVLTHAQFSPPDGLNYNHFVSKRSNALLKVIQTGAQLKKQDLQGFSIPVILVENSGRCHKNESDEKILPCGTSWIPNLFNKITEISFNGNKAIHVDKKLVEGPNPNERGKKLIPLMFAFQYLLVMKPLVRAIKSDVSRESKPAWELRDSGLASRRS rbs_tobac Ribulose bisphosphate carboxylase small subunit, chloroplastic RBCS TSSU3-8 protein protein tobac Nicotiana tabacum (Common tobacco) Common tobacco MASSVLSSAAVATRSNVAQANMVAPFTGLKSAASFPVSRKQNLDITSIASNGGRVQCMQVWPPINKKKYETLSYLPDLSQEQLLSEVEYLLKNGWVPCLEFETEHGFVYRENNKSPGYYDGRYWTMWKLPMFGCTDATQVLAEVEEAKKAYPQAWIRIIGFDNVRQVQCISFIAYKPEGY tc159_arath Translocase of chloroplast 159, chloroplastic T14P8.24 At4g02510 TOC159 PPI2 TOC160 TOC86 Translocase of chloroplast 160, chloroplastic Translocase of chloroplast 86, chloroplastic 159 kDa chloroplast outer envelope protein Plastid protein import 2 T10P11.19 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MDSKSVTPEPTNPFYASSGQSGKTYASVVAAAAAAAADKEDGGAVSSAKELDSSSEAVSGNSDKVGADDLSDSEKEKPNLVGDGKVSDEVDGSLKEDSTTPEATPKPEVVSGETIGVDDVSSLSPKPEAVSDGVGVVEENKKVKEDVEDIKDDGESKIENGSVDVDVKQASTDGESESKVKDVEEEDVGTKKDDEGESELGGKVDVDDKSDNVIEEEGVELTDKGDVIVNSSPVESVHVDVAKPGVVVVGDAEGSEELKINADAETLEVANKFDQIGDDDSGEFEPVSDKAIEEVEEKFTSESDSIADSSKLESVDTSAVEPEVVAAESGSEPKDVEKANGLEKGMTYAEVIKAASAVADNGTKEEESVLGGIVDDAEEGVKLNNKGDFVVDSSAIEAVNVDVAKPGVVVVGDVEVSEVLETDGNIPDVHNKFDPIGQGEGGEVELESDKATEEGGGKLVSEGDSMVDSSVVDSVDADINVAEPGVVVVGAAKEAVIKEDDKDDEVDKTISNIEEPDDLTAAYDGNFELAVKEISEAAKVEPDEPKVGVEVEELPVSESLKVGSVDAEEDSIPAAESQFEVRKVVEGDSAEEDENKLPVEDIVSSREFSFGGKEVDQEPSGEGVTRVDGSESEEETEEMIFGSSEAAKQFLAELEKASSGIEAHSDEANISNNMSDRIDGQIVTDSDEDVDTEDEGEEKMFDTAALAALLKAATGGGSSEGGNFTITSQDGTKLFSMDRPAGLSSSLRPLKPAAAPRANRSNIFSNSNVTMADETEINLSEEEKQKLEKLQSLRVKFLRLLQRLGHSAEDSIAAQVLYRLALLAGRQAGQLFSLDAAKKKAVESEAEGNEELIFSLNILVLGKAGVGKSATINSILGNQIASIDAFGLSTTSVREISGTVNGVKITFIDTPGLKSAAMDQSTNAKMLSSVKKVMKKCPPDIVLYVDRLDTQTRDLNNLPLLRTITASLGTSIWKNAIVTLTHAASAPPDGPSGTPLSYDVFVAQCSHIVQQSIGQAVGDLRLMNPSLMNPVSLVENHPLCRKNREGVKVLPNGQTWRSQLLLLCYSLKVLSETNSLLRPQEPLDHRKVFGFRVRSPPLPYLLSWLLQSRAHPKLPGDQGGDSVDSDIEIDDVSDSEQEDGEDDEYDQLPPFKPLRKTQLAKLSNEQRKAYFEEYDYRVKLLQKKQWREELKRMKEMKKNGKKLGESEFGYPGEEDDPENGAPAAVPVPLPDMVLPPSFDSDNSAYRYRYLEPTSQLLTRPVLDTHGWDHDCGYDGVNAEHSLALASRFPATATVQVTKDKKEFNIHLDSSVSAKHGENGSTMAGFDIQNVGKQLAYVVRGETKFKNLRKNKTTVGGSVTFLGENIATGVKLEDQIALGKRLVLVGSTGTMRSQGDSAYGANLEVRLREADFPIGQDQSSFGLSLVKWRGDLALGANLQSQVSVGRNSKIALRAGLNNKMSGQITVRTSSSDQLQIALTAILPIAMSIYKSIRPEATNDKYSMY ti110_arath Protein TIC110, chloroplastic TIC110 Translocon at the inner envelope membrane of chloroplasts 110 F4H5.1 F10K1.33 At1g06950 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MNPSLVTAINAPISPSPRSPLLSHFLPTLPHRFSKSECLSRRRYRVSFPRSSAASSDQLSVSTQAKNPGIHGNKKELTGLQPIVEKMTPPVRLATSAVVLAASLATGYGLGLRLAGSRNIAFGGAAVAGAAGGAVVYALNSAVPEVAAISLHNYVAEFEDPASVTKDDVEKIADRYGVNKGDEAFQAEICDIYCRYVTSVLPTEGQSLKGDEVAKIVKFKNALGIDEPDAAAMHMEIGRRIFRQRLETGEREGDAEQRRAFMRLVYVSALVFGDASSFLLPWKRVLKVTDAQVEIAIRENAKQLYAERLKLVGRDINVENLVDLRKSQLSFKLSDELAEDLFREHTRKVVVENISSALSILKSRTRAAKSLASVVEELEKVLEFNNLLVSLKSHSEADQFARGVGPISLIGDESDFERRMDDLKLLYRAYVTDALSGGRLEENKLVAMSQLRNILGLGKREAEAISVDVTSKSYRKRLANAVSSGDLEAQDSKAKYLQKLCEELHFDAQKAGAIHEEIYRQKLQQCVTDGELSDDNVAALLRLRVMLCIPQQTVDTAHAEICGTIFEKVVRDAISSGVDGYDAETRKSVRKAAHGLRLSRETAMSIASKAVRRVFTNYIRRARAAENRTDSAKELKKMIAFNTLVVTEMVADIKGESSDKAPEEDPVQEKEEDDEDEEWGSLESLRKTRPDKELAEKMGKPGQTEITLKDDLPDRDRIDLYKTYLLYCVTGEVTRIPFGAQITTKRDDSEYLLLNQLGGILGLSSKEIVNIHVGLAEQAFRQQAEVILADGQLTKARVEQLDELQKQVGLPQPQAEKVIKNITTTKMANAIETAVNQGRLNIKQIRELKEANVSLDSMIAVSLREKLFKKTVSDIFSSGTGEFDETEVYQTIPSDLSIDVEKAKRVVHDLAQSRLSNSLVQAVALLRQRNSKGVVLSLNDLLACDKAVPAEPMSWEVSEELSDLYAIYSKSDPKPAPEKVLRLQYLLGIDDSTATALREMEDGALSSAAEEGNFVF tc754_arath Protein TOC75-4, chloroplastic TOC75-4 F23J3.110 At4g09080 75 kDa translocon at the outer-envelope-membrane of chloroplasts 4 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEAVKEAVRKIKSLVIPHADEKDNGIVFEIKLNETDQRVEKWGLDPSLDFFEVTGNCNLGRPNSEGSNQSLMGSVTIRNIFNPKLDDLLSKIEYVRFLEAVKKPRNRTFKTSFFNSRKLSPVFTGGPGYEDLVPPMFVGRDCLKATITENLTRQRELTYGVMFEEIITRDENRRISENGLLLSPDGGISINGPPTTLSGTGIDHIATLQANITRDNTKLVNGAVVGEKNIFQVDQGLGIGNNFPLFNRHQLSLTSFIQLKQVEEGSDKPQPPVLVLHGRYGGCIGDLPSYDVFALGGPNSVRGYSMGELGAAKNILELGAEIRIPVKNTHVYAFAEHGNDLGSSKDVKGNPTGLYRKMGHGSSYGLGVKLGMVRAEYTVRHNRGTGALFLRFGERY oep37_arath Outer envelope pore protein 37, chloroplastic OEP37 Chloroplastic outer envelope pore protein of 37 kDa F6E13.8 At2g43950 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MADPSSQNPNLATPPPPSSPSPTHQIQSGTSELSPPSRPPCSTLSFLKTANRPKLRVTSEFDSDSLLFLNKVSCKLFDNLAKLKLSFQNNSQREISQPQVSFTSKHVSVLYDVEEKNTFIKSTLDVHPRLQLRALHNVKAQQGEVAMEANLTEPGYSLELSSPVPIGYPRATLKFPLGEISLQEKDEEEEEKQKRTLSVNGILKRQVMNGVCTALYTDEELRLRYAYKDDALSFIPSISLPSNAASFAFKRRFSPSDKLSYWYNFDSNMWSAVYKRTYGKDYKLKAGYDSDVRLGWASLWVGDEAGKVKTTPMKMKVQFMLQVPQDDIKSSVLMFRVKKRWDI tic40_arath Protein TIC 40, chloroplastic TIC40 PDE120 At5g16620 Protein PIGMENT DEFECTIVE EMBRYO 120 Translocon at the inner envelope membrane of chloroplasts 40 MTG13.6 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MENLTLVSCSASSPKLLIGCNFTSSLKNPTGFSRRTPNIVLRCSKISASAQSQSPSSRPENTGEIVVVKQRSKAFASIFSSSRDQQTTSVASPSVPVPPPSSSTIGSPLFWIGVGVGLSALFSYVTSNLKKYAMQTAMKTMMNQMNTQNSQFNNSGFPSGSPFPFPFPPQTSPASSPFQSQSQSSGATVDVTATKVETPPSTKPKPTPAKDIEVDKPSVVLEASKEKKEEKNYAFEDISPEETTKESPFSNYAEVSETNSPKETRLFEDVLQNGAGPANGATASEVFQSLGGGKGGPGLSVEALEKMMEDPTVQKMVYPYLPEEMRNPETFKWMLKNPQYRQQLQDMLNNMSGSGEWDKRMTDTLKNFDLNSPEVKQQFNQIGLTPEEVISKIMENPDVAMAFQNPRVQAALMECSENPMNIMKYQNDKEVMDVFNKISQLFPGMTG op24a_arath Outer envelope pore protein 24A, chloroplastic OEP24A Chloroplastic outer envelope pore protein of 24 kDa A F27F5.25 At1g45170 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MMKASFKGKFDVDKSGSVASLTFNAGNAKLRATMTDASFVAGPSFNGLSLAVEKPGFFIIDYNVPKKDVRFQFMNTIRIAEKPLNLTYIHMRGDNRTIVDGSFVIDPANKLSANYMVGTKNCKLKYTYVHGGIATFEPCYDVAKNMWDFAISHKLYGGDNLKATYQTSSKMLGLEWSNNSKSTGSFKVCASMNLAEELKPPKLTAETTWNLEL gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 toc34-rbcs-2 Arabidopsis Toc34 interacts with Tobacco RBCS 733276 N/A 733284 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) N/A 733285 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro direct interaction direct interaction false false false Figure 3B GMP-PNP, a non-hydrolyzable analog of GTP, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. Radiolabelling of P69249 was carried out using wheat germ lysate. toc34-rbcs-3 Arabidopsis Toc34 interacts with Tobacco RBCS 733276 N/A 733285 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 733284 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 3B GMP-PNP, a non-hydrolyzable analog of GTP, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. Phosphorylation of P69249 toc159-rbcs-1 Arabidopsis Toc159 interacts with Tobacco RBCS 733276 N/A 733285 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 733286 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 3B GMP-PNP, a non-hydrolyzable analog of GTP, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. Radiolabelling of P69249 was carried ot using reticulocyte lysate. toc34-rbcs-4 Arabidopsis Toc34 interacts with Tobacco RBCS 733276 N/A 733285 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 733284 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 3B GMP-PNP, a non-hydrolyzable analog of GTP, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. Radiolabelling of P69249 was carried out using reticulocyte lysate. toc159-toc110 Toc159 interacts with Toc110, Toc75, Tic40 and Oep37 733277 N/A 733287 unspecified role unspecified role prey prey N/A 733288 unspecified role unspecified role prey prey N/A 733289 unspecified role unspecified role prey prey N/A 733286 unspecified role unspecified role bait bait N/A 733290 unspecified role unspecified role prey prey association association false false false Figure 1C toc159-toc75-1 Toc159 interacts with Toc75, Tic34 and Oep24 733277 N/A 733291 unspecified role unspecified role prey prey N/A 733284 unspecified role unspecified role prey prey N/A 733288 unspecified role unspecified role prey prey N/A 733286 unspecified role unspecified role bait bait association association false false false Figure 6A The authors note that "incubation with GDP leads to a reduced coimmunoprecipitation of Toc159 by Toc34 antiserum and vice versa". The accession number for Oep24 (Q1H5C9) was derived from the closest amino acid sequence match for an Arabidopsis protein to the Oep24 Pisum sativaum (Pea) protein in the UniProt database. The Oep24 Pea protein was cited in a secondary reference from a reference provided by the authors. The authors concluded that "although small amounts of Oep24 were coimmunoprecipitated by the used Toc34 antiserum, it presents only a small portion of total Oep24 content in OEVs. Furthermore, Toc75, Toc159 and Toc34 were not coimmunoprecipitated by Oep24 antibodies regardless of the added nucleotides. Therefore, we consider the precipitation of Oep24 as an unspecific reaction". toc34-toc75-2 Toc34 interacts with Toc75, Tic159 and Oep24 733277 N/A 733291 unspecified role unspecified role prey prey N/A 733288 unspecified role unspecified role prey prey N/A 733286 unspecified role unspecified role prey prey N/A 733284 unspecified role unspecified role bait bait association association false false false Figure 6A The authors note that "incubation with GDP leads to a reduced coimmunoprecipitation of Toc159 by Toc34 antiserum and vice versa". The accession number for Oep24 (Q1H5C9) was derived from the closest amino acid sequence match for an Arabidopsis protein to the Oep24 Pisum sativaum (Pea) protein in the UniProt database. The Oep24 Pea protein was cited in a secondary reference from a reference provided by the authors. The authors concluded that "although small amounts of Oep24 were coimmunoprecipitated by the used Toc34 antiserum, it presents only a small portion of total Oep24 content in OEVs. Furthermore, Toc75, Toc159 and Toc34 were not coimmunoprecipitated by Oep24 antibodies regardless of the added nucleotides. Therefore, we consider the precipitation of Oep24 as an unspecific reaction". toc159-rbcs-3 Arabidopsis Toc159 interacts with Tobacco RBCS 733278 N/A 733285 unspecified role unspecified role bait bait n-terminal 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 733286 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 4 GDP toc159-rbcs-4 Arabidopsis Toc159 interacts with Tobacco RBCS 733278 N/A 733285 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 chemical synthesis Chemical synthesis (Chemical synthesis) N/A 733286 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 4 GDP GMP-PNP, a non-hydrolyzable analog of GTP. GMP-PNP is a compound, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. toc34-rbcs-5 Arabidopsis Toc34 interacts with Tobacco RBCS 733278 N/A 733285 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 n-terminal 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 733284 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 4 GMP-PNP, a non-hydrolyzable analog of GTP. GMP-PNP is a compound, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. toc34-rbcs Arabidopsis Toc34 interacts with Tobacco RBCS 733279 N/A 733284 unspecified role unspecified role bait bait arath-leaf Arabidopsis thaliana leaf leaf leaf N/A 733285 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro direct interaction direct interaction false false false Figure 3A GMP-PNP, a non-hydrolyzable analog of GTP. GMP-PNP is a compound in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. toc159-rbcs Arabidopsis Toc159 interacts with Tobacco RBCS 733279 N/A 733285 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 733286 unspecified role unspecified role bait bait arath-leaf Arabidopsis thaliana leaf leaf leaf direct interaction direct interaction false false false Figure 3A GMP-PNP, a non-hydrolyzable analog of GTP. GMP-PNP is a compound, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. toc159-rbcs-2 Arabidopsis Toc159 interacts with Tobacco RBCS 733280 N/A 733285 unspecified role unspecified role bait bait in vitro In vitro N/A 733286 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 3C GMP-PNP, a non-hydrolyzable analog of GTP, in which the oxygen atom bridging the beta to the gamma phosphate is replaced by a nitrogen atom. It binds tightly to G-protein in the presence of Mg2+. The nucleotide is a potent stimulator of adenylate cyclase. GTP GDP toc34-toc159 Arabidopsis Toc34 interacts with Toc159 733280 N/A 733284 unspecified role unspecified role bait bait c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 733286 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 6E Presence of chemically synthesised peptides covering 1 to 21 and 22 to 47 amino acid residues of the pSSU protein (P69249). The authors note that "In contrast, Toc159GDP" Toc159 pretreated with GDP "shows reduced binding to Toc34". toc159-rbcs-6 Arabidopsis Toc159 is a GTPase 733281 N/A 733286 enzyme enzyme neutral component neutral component ecoli Escherichia coli (strain K12) N/A 733292 enzyme target enzyme target substrate substrate neutral component neutral component gtpase reaction gtpase reaction false false false Figure 5B A chemically synthesised pSSU peptide (region 22 to 47 amino acid residues) was found to stimulate GTPase activity of Toc159f by about four-fold. toc34-rbcs-7 Arabidopsis Toc34 is a GTPase 733281 N/A 733284 enzyme enzyme neutral component neutral component N/A 733292 enzyme target enzyme target substrate substrate neutral component neutral component gtpase reaction gtpase reaction false false false Figure 5C toc159-toc75-2 Toc159 interacts with Toc75 733282 N/A 733286 unspecified role unspecified role neutral component neutral component N/A 733288 unspecified role unspecified role neutral component neutral component N/A 733284 unspecified role unspecified role neutral component neutral component physical association physical association false false false Figure 6C The authors note that "in the presence of GMP-PNP, the Toc components are enriched in higher density fractions as compared to the absence of nucleotide". toc159-toc75-3 Toc159 interacts with Toc75 733283 N/A 733288 unspecified role unspecified role neutral component neutral component N/A 733286 unspecified role unspecified role neutral component neutral component N/A 733284 unspecified role unspecified role neutral component neutral component physical association physical association false false false Figure 6D The authors note that "in the presence of GMP-PNP, the Toc components are enriched in higher density fractions as compared to the absence of nucleotide" amnd that "In the presence of GDP, no Toc components were found in a fraction of high sucrose concentrations". The authors conclude that "GMP-PNP leads to a stabilisation of the Toc complex, whereas GDP weakens the interaction between the Toc components".