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/ A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. Cell (0092-8674) 2009 imex curation Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. hbellen@bcm.tmc.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 12 in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel anti tag western anti tag western blot anti tag western Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. Homo-multimers were formed both in detergent-solubilized phospholipid liposomes hbellen@bcm.tmc.edu Accepted 2009-Not a month!-30 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 12 imex curation A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. Cell (0092-8674) 2009 imex curation Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. hbellen@bcm.tmc.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 12 in vitro In vitro crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. Interaction observed using different cross-linking agents Bis(Sulfosuccinimidyl) suberate (BS3), disuccinimidyl suberate (DSS), 1,5-Difluoro-2,4-dinitrobenzene (DFDNB), or Dithiobis(succinimidyl propionate) (DSP). hbellen@bcm.tmc.edu Accepted 2009-Not a month!-30 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 12 imex curation A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. Cell (0092-8674) 2009 imex curation Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. hbellen@bcm.tmc.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 12 drome Drosophila melanogaster (Fruit fly) Fruit fly anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. hbellen@bcm.tmc.edu Accepted 2009-Not a month!-30 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 12 imex curation A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. A synaptic vesicle-associated Ca2+ channel promotes endocytosis and couples exocytosis to endocytosis. Cell (0092-8674) 2009 imex curation Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. hbellen@bcm.tmc.edu 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 12 drome-head Drosophila melanogaster head anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Yao CK., Lin YQ., Ly CV., Ohyama T., Haueter CM., Moiseenkova-Bell VY., Wensel TG., Bellen HJ. Existence of higher molecular weight homo-oligomers were identified by the molecular weight of anti-tag blots. hbellen@bcm.tmc.edu Accepted 2009-Not a month!-30 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Cell (0092-8674) 2009 jyoti Only protein-protein interactions 12 imex curation flowr_drome Calcium channel flower fwe CG6151 flower protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSFAEKITGLLARPNQQDPIGPEQPWYLKYGSRLLGIVAAFFAILFGLWNVFSIITLSVSCLVAGILQMVAGFVVMLLEAPCCFVCFGQVNEIAEKVESKPLYFRAGLYIAMAIPPIILCFGLASLFGSGLIFGTGVVYGMMALGKKASAEDMRAAAQQTFGGNTPAQTNDRAGIVNNAQPFSFTGAVGTDSNV 2D2817158AD73A57 q95t12-2 Calcium channel flower fwe CG6151 flower protein protein drome Drosophila melanogaster (Fruit fly) Fruit fly MSFAEKITGLLARPNQQDPIGPEQPWYLKYGSRLLGIVAAFFAILFGLWNVFSIITLSVSCLVAGILQMVAGFVVMLLEAPCCFVCFGQVNEIAEKVESKPLYFRAGLYIAMAIPPIILCFGLASLFGSGLIFGTGVVYGMMALGKKASREDMAAAATSPTQMAGSQAGGQMQMGGDQHITLMEDPDVWRPT C305833C7292F2E3 flower-1 3 6 unspecified role unspecified role prey prey n-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association CoIP of PB-HA with anti-HA antibody allows detection of the PB-GFP protein by western blotting, and vice versa (Figure 5B), suggesting that Flower has the ability to form at least a homodimer. Fig. 5B flower-2 3 6 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait n-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association CoIP of PB-HA with anti-HA antibody allows detection of the PB-GFP protein by western blotting, and vice versa (Figure 5B), suggesting that Flower has the ability to form at least a homodimer. Fig. 5B flower-3 4 6 unspecified role unspecified role neutral component neutral component n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association To test whether Flower can form higher multimers, the PB-HA proteins from head extracts of PB-HA-rescued adult flies were immunoprecipitated with anti-HA antibody. Fig. 5C Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-4 4 6 unspecified role unspecified role neutral component neutral component n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association To test whether Flower can form higher multimers, the PB-HA proteins from head extracts of PB-HA-rescued adult flies were immunoprecipitated with anti-HA antibody. Fig. 5C Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-5 4 6 unspecified role unspecified role neutral component neutral component n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association To test whether Flower can form higher multimers, the PB-HA proteins from head extracts of PB-HA-rescued adult flies were immunoprecipitated with anti-HA antibody. Fig. 5C Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-6 1 5 unspecified role unspecified role neutral component neutral component c-terminal tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Purified Flower-1D4 proteins or Flower-containing proteoliposomes (PL) were incubated with 0.5, 1, or 4% perfluoro-octanoic acid (PFO) sample buffers and run on a 4%�12% gradient Tris/glycine precast native gel. Fig. 7B, Supp Fig. 6 Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-7 1 5 unspecified role unspecified role neutral component neutral component c-terminal tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Purified Flower-1D4 proteins or Flower-containing proteoliposomes (PL) were incubated with 0.5, 1, or 4% perfluoro-octanoic acid (PFO) sample buffers and run on a 4%�12% gradient Tris/glycine precast native gel. Fig. 7B, Supp. Fig. 6 Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-8 1 5 unspecified role unspecified role neutral component neutral component c-terminal tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Purified Flower-1D4 proteins or Flower-containing proteoliposomes (PL) were incubated with 0.5, 1, or 4% perfluoro-octanoic acid (PFO) sample buffers and run on a 4%�12% gradient Tris/glycine precast native gel. Fig. 7B, Supp. Fig. 6 Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-9 2 5 unspecified role unspecified role neutral component neutral component c-terminal 1D4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast direct interaction direct interaction Purified Flower-1D4 proteins or Flower-containing proteoliposomes (PL) were incubated with 0.5, 1, or 4% perfluoro-octanoic acid (PFO) sample buffers and run on a 4%�12% gradient Tris/glycine precast native gel. Fig. 7B Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-10 2 5 unspecified role unspecified role neutral component neutral component c-terminal 1D4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast direct interaction direct interaction When 15 ng purified Flower-1D4 protein was incubated at RT for 5 min with 2% SDS, homo-multimers were observed on western blots with the antibody against the 1D4 peptide. Supp. Fig. 6 Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-11 2 5 unspecified role unspecified role neutral component neutral component c-terminal 1D4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast direct interaction direct interaction When 15 ng purified Flower-1D4 protein was incubated at RT for 5 min with 2% SDS, homo-multimers were observed on western blots with the antibody against the 1D4 peptide. Supp. Fig. 6 Homodimers, homotrimers, homotetramers and higher order multimers were observed. flower-12 2 5 unspecified role unspecified role neutral component neutral component c-terminal 1D4 tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast direct interaction direct interaction When 15 ng purified Flower-1D4 protein was incubated at RT for 5 min with 2% SDS, homo-multimers were observed on western blots with the antibody against the 1D4 peptide. Supp. Fig. 6 Homodimers, homotrimers, homotetramers and higher order multimers were observed.