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/ The plastid division protein AtMinD1 is a Ca2+-ATPase stimulated by AtMinE1. The plastid division protein AtMinD1 is a Ca2+-ATPase stimulated by AtMinE1. J. Biol. Chem. (0021-9258) 2005 imex curation Aldridge C., Moller SG. His-AtMinD1 fusion protein was expressed in E. coli followed by denaturing Co2+ affinity chromatography purification and refolding by dialysis sgm5@le.ac.uk 03-Mar-2009: Contacted by JYOTI. 03-Feb-2009: Contacted by JYOTI. orchard Only protein-protein interactions tobac-leaf Nicotiana tabacum (tobacco) leaf leaf leaf bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence Aldridge C., Moller SG. His-AtMinD1 fusion protein was expressed in E. coli followed by denaturing Co2+ affinity chromatography purification and refolding by dialysis sgm5@le.ac.uk Accepted 2008-Not a month!-3 by ORCHARD. 03-Mar-2009: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation The plastid division protein AtMinD1 is a Ca2+-ATPase stimulated by AtMinE1. The plastid division protein AtMinD1 is a Ca2+-ATPase stimulated by AtMinE1. J. Biol. Chem. (0021-9258) 2005 imex curation Aldridge C., Moller SG. His-AtMinD1 fusion protein was expressed in E. coli followed by denaturing Co2+ affinity chromatography purification and refolding by dialysis sgm5@le.ac.uk 03-Mar-2009: Contacted by JYOTI. 03-Feb-2009: Contacted by JYOTI. orchard Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Aldridge C., Moller SG. His-AtMinD1 fusion protein was expressed in E. coli followed by denaturing Co2+ affinity chromatography purification and refolding by dialysis sgm5@le.ac.uk Accepted 2008-Not a month!-3 by ORCHARD. 03-Mar-2009: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation The plastid division protein AtMinD1 is a Ca2+-ATPase stimulated by AtMinE1. The plastid division protein AtMinD1 is a Ca2+-ATPase stimulated by AtMinE1. J. Biol. Chem. (0021-9258) 2005 imex curation Aldridge C., Moller SG. His-AtMinD1 fusion protein was expressed in E. coli followed by denaturing Co2+ affinity chromatography purification and refolding by dialysis sgm5@le.ac.uk 03-Mar-2009: Contacted by JYOTI. 03-Feb-2009: Contacted by JYOTI. orchard Only protein-protein interactions in vitro In vitro atpase assay atpase assay autoradiography autoradiography Aldridge C., Moller SG. His-AtMinD1 fusion protein was expressed in E. coli followed by denaturing Co2+ affinity chromatography purification and refolding by dialysis sgm5@le.ac.uk Accepted 2008-Not a month!-3 by ORCHARD. 03-Mar-2009: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2005 orchard Only protein-protein interactions imex curation mind1_arath Putative septum site-determining protein minD homolog, chloroplastic MIND1 At5g24020 ARC11 Protein ACCUMULATION AND REPLICATION OF CHLOROPLASTS 11 Septum site-determining protein MinD1 MZF18.10 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MASLRLFSTNHQSLLLPSSLSQKTLISSPRFVNNPSRRSPIRSVLQFNRKPELAGETPRIVVITSGKGGVGKTTTTANVGLSLARYGFSVVAIDADLGLRNLDLLLGLENRVNYTCVEVINGDCRLDQALVRDKRWSNFELLCISKPRSKLPMGFGGKALEWLVDALKTRPEGSPDFIIIDCPAGIDAGFITAITPANEAVLVTTPDITALRDADRVTGLLECDGIRDIKMIVNRVRTDMIKGEDMMSVLDVQEMLGLSLLGVIPEDSEVIRSTNRGFPLVLNKPPTLAGLAFEQAAWRLVEQDSMKAVMVEEEPKKRGFFSFFGG BE7AEF4307167825 mine1_arath Cell division topological specificity factor homolog, chloroplastic MINE1 ARC12 F10D13.22 At1g69390 Protein ACCUMULATION AND REPLICATION OF CHLOROPLASTS 12 F23O10.25 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAMSSGTLRISATLVSPYHHHHRNRLSLPSSSSKVDFTGFISNGVNSLETQKCTPGLAISRENTRGQVKVLARNTGDYELSPSPAEQEIESFLYNAINMGFFDRLNLAWKIIFPSHASRRSSNARIAKQRLKMILFSDRCDVSDEAKRKIVNNIIHALSDFVEIESEEKVQLNVSTDGDLGTIYSVTVPVRRVKPEYQDVDEAGTITNVEYKDTRDGSVDVRFDFYVPE FC307A9E3C4DE123 atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mind-atp 3 6 enzyme target enzyme target substrate substrate neutral component neutral component 4 enzyme enzyme 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 Q9MBA2:p.Lys72Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain K A Wed May 02 18:39:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys72ala' Tue Jun 30 17:16:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys72Ala' ecoli Escherichia coli (strain K12) atpase reaction atpase reaction Figure 2 Characterization of AtMinD1 ATP hydrolysis. A, increasing amounts of AtMinD1 lead to a linear increase in ATP hydrolysis. The authors concluded that AtMinD1 ATPase activity and that AtMinD1 ATP hydrolysis is mediated through the Walker A (GXXXXGKT/S) domain. Km = 500 micromole ATP. Vmax = 0.2 pmol ATP per second pH 7.4 Temp = 35C AtMinE1 had a marked effect on the amount of Pi release, showing an ~3-fold increase in ATP hydrolysis compared with AtMinD1 alone and taking into account background Pi release in assays only containing AtMinE1 Note that EcMinE (MinE of E.coli origin) was unable to stimulate AtMinD1 activity mind-mine 2 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait Q9MBA2:p.Lys72Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:25:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys72ala' Tue Jun 30 19:11:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys72Ala' physical association physical association Figure 5A AtMinD1(K72A) Is Unable to Interact with AtMinE1 mind-mine-1 2 5 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey Q9MBA2:p.Lys72Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 18:01:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys72ala' Tue Jun 30 17:36:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys72Ala' physical association physical association Figure 5A AtMinD1(K72A) Is Unable to Interact with AtMinE1 mind-3 2 4 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey Q9MBA2:p.Lys72Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:25:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys72ala' Tue Jun 30 15:49:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys72Ala' physical association physical association Figure 5A mind-2 2 4 unspecified role unspecified role bait bait 4 unspecified role unspecified role prey prey Q9MBA2:p.Ala296Gly mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain A G Wed May 02 18:21:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala296gly' Tue Jun 30 18:12:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala296Gly' physical association physical association Figure 5A mind-1 1 4 unspecified role unspecified role neutral component neutral component region C-terminal YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role neutral component neutral component region N-terminal YFP protein yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Figure 5B tobacco co-bombarded with pWEN-NY/AtMinD1 and pWEN-CY/AtMinD1 showed clear fluorescence (Fig. 5B). Tobacco co-bombarded with pWEN-NY/AtMinD1(K72A) and pWEN-CY/AtMinD1 also showed clear fluorescence (Fig. 5B), demonstrating that AtMinD1(K72A) is able to dimerize in planta and showing that AtMinD1(K72A) mislocalization is not because of loss of dimerization as observed for ARC11/AtMinD1 (A296G). Tobacco cells bombarded with single vectors (negative controls) showed no fluorescence, as expected.