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/ reddy-1999-1 Interaction of a kinesin-like protein with calmodulin isoforms from Arabidopsis. in vitro In vitro biochemical biochemical weight by comassie molecular weight estimation by coomasie staining weight by comassie Reddy VS., Safadi F., Zielinski RE., Reddy AS. It was not possible to unambiguously identify the bovine calmodulin protein used in this study. Therefore all interactions involving this protein were not curated. The interaction of the AtCaM isoforms with the Arabidopsis KCBP synthetic peptide (CaM-binding region of KCBP) was analyzed by electrophoretic mobility shift assay in urea containing gels. Each of the AtCaM isoforms and the bovine CaM (178 pmol) was incubated with KCBP synthetic peptide (712 pmol) in the presence of 4M urea, 100 mM Tris-HCl, pH 7.5, and 1 mM CaCl2 or 5 mM EGTA at room temperature for 1 h in a total volume of 20 ml. Then 10 ml of sample buffer (0.375 M Tris-HCl, pH 6.8, 30 percent glycerol, and 0.023 percent bromphenol blue) was added to the samples and electrophoresed in 12 percent polyacrylamide gels containing 4 M urea, 0.375 M Tris, pH 8.8, and either 1 mM CaCl2 or 5 mM EGTA. The gels were run at a constant voltage of 25 V/gel in an electrode buffer (25 mM Tris-HCl, pH 8.3, 192mM glycine, and either 1 mM CaCl2 or 5 mM EGTA). The gels were stained with 0.25 percent Coomassie Blue R-250 in 7.5 percent acetic acid and 50 percent methanol for 1 h and then destained with 30 percent methanol and 7 percent acetic acid. reddy@lamar.colostate.edu J. Biol. Chem. (0021-9258) 1999 Only protein-protein interactions imex curation reddy-1999-2 Interaction of a kinesin-like protein with calmodulin isoforms from Arabidopsis. in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay predetermined predetermined participant predetermined Reddy VS., Safadi F., Zielinski RE., Reddy AS. It was not possible to unambiguously identify the bovine calmodulin protein used in this study. Therefore all interactions involving this protein were not curated. reddy@lamar.colostate.edu J. Biol. Chem. (0021-9258) 1999 Only protein-protein interactions imex curation The interaction of the AtCaM isoforms with the Arabidopsis KCBP synthetic peptide (CaM-binding region of KCBP) was analyzed by electrophoretic mobility shift assay in urea containing gels. Each of the AtCaM isoforms and the bovine CaM (178 pmol) was incubated with KCBP synthetic peptide (712 pmol) in the presence of 4M urea, 100 mM Tris-HCl, pH 7.5, and 1 mM CaCl2 or 5 mM EGTA at room temperature for 1 h in a total volume of 20 ml. Then 10 ml of sample buffer (0.375 M Tris-HCl, pH 6.8, 30 percent glycerol, and 0.023 percent bromphenol blue) was added to the samples and electrophoresed in 12 percent polyacrylamide gels containing 4 M urea, 0.375 M Tris, pH 8.8, and either 1 mM CaCl2 or 5 mM EGTA. The gels were run at a constant voltage of 25 V/gel in an electrode buffer (25 mM Tris-HCl, pH 8.3, 192mM glycine, and either 1 mM CaCl2 or 5 mM EGTA). The gels were stained with 0.25 percent Coomassie Blue R-250 in 7.5 percent acetic acid and 50 percent methanol for 1 h and then destained with 30 percent methanol and 7 percent acetic acid. calm6_arath Calmodulin-6 CAM6 F13M11 At5g21274 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MADQLTDDQISEFKEAFSLFDKDGDGCITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLNLMARKMKDTDSEEELKEAFRVFDKDQNGFISAAELRHVMTNLGEKLSDEEVDEMIREADVDGDGQINYEEFVKVMMAK kn14e_arath Kinesin-like protein KIN-14E KIN14E KCBP Kinesin-like calmodulin-binding protein Protein ZWICHEL ZWI K14B20.10 At5g65930 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEGQRGSNSSLSSGNGTEVATDVSSCFYVPNPSGTDFDAESSSLPPLSPAPQVALSIPAELAAAIPLIDRFQVEAFLRLMQKQIQSAGKRGFFYSKKSSGSNVRERFTFEDMLCFQKDPIPTSLLKINSDLVSRATKLFHLILKYMGVDSSDRSTPPSLDERIDLVGKLFKKTLKRVELRDELFAQISKQTRHNPDRQYLIKAWELMYLCASSMPPSKDIGGYLSEYIHNVAHDATIEPDAQVLAVNTLKALKRSIKAGPRHTTPGREEIEALLTGRKLTTIVFFLDETFEEISYDMATTVSDAVEELAGTIKLSAFSSFSLFECRKVVSSSKSSDPGNEEYIGLDDNKYIGDLLAEFKAIKDRNKGEILHCKLVFKKKLFRESDEAVTDLMFVQLSYVQLQHDYLLGNYPVGRDDAAQLCALQILVGIGFVNSPESCIDWTSLLERFLPRQIAITRAKREWELDILARYRSMENVTKDDARQQFLRILKALPYGNSVFFSVRKIDDPIGLLPGRIILGINKRGVHFFRPVPKEYLHSAELRDIMQFGSSNTAVFFKMRVAGVLHIFQFETKQGEEICVALQTHINDVMLRRYSKARSAANSLVNGDISCSSKPQNFEVYEKRLQDLSKAYEESQKKIEKLMDEQQEKNQQEVTLREELEAIHNGLELERRKLLEVTLDRDKLRSLCDEKGTTIQSLMSELRGMEARLAKSGNTKSSKETKSELAEMNNQILYKIQKELEVRNKELHVAVDNSKRLLSENKILEQNLNIEKKKKEEVEIHQKRYEQEKKVLKLRVSELENKLEVLAQDLDSAESTIESKNSDMLLLQNNLKELEELREMKEDIDRKNEQTAAILKMQGAQLAELEILYKEEQVLRKRYYNTIEDMKGKIRVYCRIRPLNEKESSEREKQMLTTVDEFTVEHPWKDDKRKQHIYDRVFDMRASQDDIFEDTKYLVQSAVDGYNVCIFAYGQTGSGKTFTIYGHESNPGLTPRATKELFNILKRDSKRFSFSLKAYMVELYQDTLVDLLLPKSARRLKLEIKKDSKGMVFVENVTTIPISTLEELRMILERGSERRHVSGTNMNEESSRSHLILSVVIESIDLQTQSAARGKLSFVDLAGSERVKKSGSAGCQLKEAQSINKSLSALGDVIGALSSGNQHIPYRNHKLTMLMSDSLGGNAKTLMFVNVSPAESNLDETYNSLLYASRVRTIVNDPSKHISSKEMVRLKKLVAYWKEQAGKKGEEEDLVDIEEDRTRKDEADS calm5_arath Calmodulin-5 CAM5 TCH1 T20P8.8 At2g27030 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MADQLTDDQISEFKEAFSLFDKDGDGCITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLNLMARKMKDTDSEEELKEAFRVFDKDQNGFISAAELRHVMTNLGEKLTDEEVDEMIKEADVDGDGQINYEEFVKVMMAK calm1_arath Calmodulin-1/4 CAM1 At5g37780 K22F20.2 T31G3.3 CAM4 At1g66410 T27F4.1 F28G11.13 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MADQLTDEQISEFKEAFSLFDKDGDGCITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLNLMAKKMKDTDSEEELKEAFRVFDKDQNGFISAAELRHVMTNLGEKLTDEEVEEMIREADVDGDGQINYEEFVKIMMAK q39130-cam6 Q39130 interacts with Cam6 730659 N/A 730661 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) N/A 730662 unspecified role unspecified role neutral component neutral component 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) direct interaction direct interaction false false false Figure 1 The authors state that the interaction is dependent on the presence of Calcium ions (but do not present this data). q39130-cam2 Q39130 interacts with Cam2 730659 N/A 730663 unspecified role unspecified role neutral component neutral component N/A 730662 unspecified role unspecified role neutral component neutral component 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) direct interaction direct interaction false false false Figure 1 The authors state that the interaction is dependent on the presence of Calcium ions (but do not present this data). q39130-cam4 Q39130 interacts with Cam4 730659 N/A 730664 unspecified role unspecified role neutral component neutral component N/A 730662 unspecified role unspecified role neutral component neutral component 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) direct interaction direct interaction false false false Figure 1 The authors state that the interaction is dependent on the presence of Calcium ions (but do not present this data). q39130-cam6-1 Q39130 interacts with Cam6 730660 N/A 730661 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 730662 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Figure 4C The authors state the interaction was dependent on the presence of calcium ions (but do not present data to substantiate this). Dissociation constant was 49 +/- 1.2nm q39130-cam2-1 Q39130 interacts with Cam2 730660 N/A 730663 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 730662 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Figure 4A The authors state that the interaction is dependent on the presence of Calcium ions (but do not present this data). Dissociation constant was 20 +/- 0.04 nm. q39130-cam4-1 Q39130 interacts with Cam4 730660 N/A 730662 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 ecobb Escherichia coli (strain B / BL21) N/A 730664 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 4B The authors state that the interaction is dependent on the presence of Calcium ions (but do not present this data). Dissociation constant was 49 +/- 0.96 nm.