bhf-ucl Mutant Frizzled 4 associated with vitreoretinopathy traps wild-type Frizzled in the endoplasmic reticulum by oligomerization. Mutant Frizzled 4 associated with vitreoretinopathy traps wild-type Frizzled in the endoplasmic reticulum by oligomerization. Nat. Cell Biol. (1465-7392) 2004 imex curation Kaykas A., Yang-Snyder J., Héroux M., Shah K.V., Bouvier M., Moon R.T. Cardiac - Interactions involving cardiac related proteins A beta-galactosidase complementation assay was used, where the two proteins of interest were each fused to the α (alpha) and ω (omega: from greek) complementary parts of beta-galactosidase. The α is a short N-terminal fragment and the ω is a long C-terminal fragment. To quentify the levels of protein expression, ω fusion proteins were fused to renilla luciferase (RL). The triple fusions allowed simultaneous measurements of beta-gal complementation versus expression levels, as measured by RL activity. rtmoon@u.washington.edu 28-Oct_2013: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Kaykas A., Yang-Snyder J., Héroux M., Shah K.V., Bouvier M., Moon R.T. Cardiac - Interactions involving cardiac related proteins Repeat of the experiment in the presence of cell-impermeable crosslinker BS suggests that Fzs interact in the plasma membrane. A beta-galactosidase complementation assay was used, where the two proteins of interest were each fused to the α (alpha) and ω (omega: from greek) complementary parts of beta-galactosidase. The α is a short N-terminal fragment and the ω is a long C-terminal fragment. To quentify the levels of protein expression, ω fusion proteins were fused to renilla luciferase (RL). The triple fusions allowed simultaneous measurements of beta-gal complementation versus expression levels, as measured by RL activity. rtmoon@u.washington.edu Accepted 2013-OCT-21 by ORCHARD 28-Oct_2013: Contacted by IntAct-Help. Nat. Cell Biol. (1465-7392) 2004 imex curation Mutant Frizzled 4 associated with vitreoretinopathy traps wild-type Frizzled in the endoplasmic reticulum by oligomerization. Mutant Frizzled 4 associated with vitreoretinopathy traps wild-type Frizzled in the endoplasmic reticulum by oligomerization. Nat. Cell Biol. (1465-7392) 2004 imex curation Kaykas A., Yang-Snyder J., Héroux M., Shah K.V., Bouvier M., Moon R.T. Cardiac - Interactions involving cardiac related proteins A beta-galactosidase complementation assay was used, where the two proteins of interest were each fused to the α (alpha) and ω (omega: from greek) complementary parts of beta-galactosidase. The α is a short N-terminal fragment and the ω is a long C-terminal fragment. To quentify the levels of protein expression, ω fusion proteins were fused to renilla luciferase (RL). The triple fusions allowed simultaneous measurements of beta-gal complementation versus expression levels, as measured by RL activity. rtmoon@u.washington.edu 28-Oct_2013: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. beta galactosidase beta galactosidase complementation beta galactosidase predetermined predetermined participant predetermined Kaykas A., Yang-Snyder J., Héroux M., Shah K.V., Bouvier M., Moon R.T. Cardiac - Interactions involving cardiac related proteins A beta-galactosidase complementation assay was used, where the two proteins of interest were each fused to the α (alpha) and ω (omega: from greek) complementary parts of beta-galactosidase. The α is a short N-terminal fragment and the ω is a long C-terminal fragment. To quentify the levels of protein expression, ω fusion proteins were fused to renilla luciferase (RL). The triple fusions allowed simultaneous measurements of beta-gal complementation versus expression levels, as measured by RL activity. rtmoon@u.washington.edu Accepted 2013-OCT-21 by ORCHARD 28-Oct_2013: Contacted by IntAct-Help. Nat. Cell Biol. (1465-7392) 2004 imex curation Mutant Frizzled 4 associated with vitreoretinopathy traps wild-type Frizzled in the endoplasmic reticulum by oligomerization. Mutant Frizzled 4 associated with vitreoretinopathy traps wild-type Frizzled in the endoplasmic reticulum by oligomerization. Nat. Cell Biol. (1465-7392) 2004 imex curation Kaykas A., Yang-Snyder J., Héroux M., Shah K.V., Bouvier M., Moon R.T. Cardiac - Interactions involving cardiac related proteins A beta-galactosidase complementation assay was used, where the two proteins of interest were each fused to the α (alpha) and ω (omega: from greek) complementary parts of beta-galactosidase. The α is a short N-terminal fragment and the ω is a long C-terminal fragment. To quentify the levels of protein expression, ω fusion proteins were fused to renilla luciferase (RL). The triple fusions allowed simultaneous measurements of beta-gal complementation versus expression levels, as measured by RL activity. rtmoon@u.washington.edu 28-Oct_2013: Contacted by IntAct-Help. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. bret bioluminescence resonance energy transfer BRET LRET LRET BRET bret tag luciferase tag visualisation by luciferase assay tag luciferase Kaykas A., Yang-Snyder J., Héroux M., Shah K.V., Bouvier M., Moon R.T. Cardiac - Interactions involving cardiac related proteins A beta-galactosidase complementation assay was used, where the two proteins of interest were each fused to the α (alpha) and ω (omega: from greek) complementary parts of beta-galactosidase. The α is a short N-terminal fragment and the ω is a long C-terminal fragment. To quentify the levels of protein expression, ω fusion proteins were fused to renilla luciferase (RL). The triple fusions allowed simultaneous measurements of beta-gal complementation versus expression levels, as measured by RL activity. rtmoon@u.washington.edu Accepted 2013-OCT-21 by ORCHARD 28-Oct_2013: Contacted by IntAct-Help. Nat. Cell Biol. (1465-7392) 2004 imex curation fzd2_rat Frizzled-2 Fzd2 protein protein rat Rattus norvegicus (Rat) Rat MRARSALPRSALPRLLLPLLLLPAAGPAQFHGEKGISIPDHGFCQPISIPLCTDIAYNQTIMPNLLGHTNQEDAGLEVHQFYPLVKVQCSPELRFFLCSMYAPVCTVLEQAIPPCRSICERARQGCEALMNKFGFQWPERLRCEHFPRHGAEQICVGQNHSEDGTPALLTTAPPSGLQPGAGGTPGGPGGGGAPPRYATLEHPFHCPRVLKVPSYLSYKFLGERDCAAPCEPARPDGSMFFSHHHTRFARLWILTWSVLCCASTFFTVTTSLVAMQRFRYPERPIIFLSGCYTMVSVAYIAGFVLQERVVCNERFSEDGYRTVGQGTKKEGCTILFMMLYFFSMASSIWWVILSLTWFLAAGMKWGHAAIEANSQYFHLAAWAVPAVKTITILAMGQIDGDLLSGVCFVGLNRLDPLRGFVLAPLFVYLFIGTSFLLAGFVSLFRIRTIMKHDGTKTEPLERLMVRIGVFSVLYTVPATIVIACYFYEQAFREHWERSWVSQHCKSLAIPCPAHYTPRTSPDFTVYMIKYLMTLIVGITSGFWIWSGKTLHSWRKFYTRLTNSRHGETTV mint Receptor for Wnt proteins. Most of frizzled receptors are coupled to the beta-catenin canonical signaling pathway, which leads to the activation of disheveled proteins, inhibition of GSK-3 kinase, nuclear accumulation of beta-catenin and activation of Wnt target genes. A second signaling pathway involving PKC and calcium fluxes has been seen for some family members, but it is not yet clear if it represents a distinct pathway or if it can be integrated in the canonical pathway, as PKC seems to be required for Wnt-mediated inactivation of GSK-3 kinase. Both pathways seem to involve interactions with G-proteins. May be involved in transduction and intercellular transmission of polarity information during tissue morphogenesis and/or in differentiated tissues. Activation by Wnt5A stimulates PKC activity via a G-protein-dependent mechanism 4FB895D9BEAFCA4E fzd4_human Frizzled-4 FZD4 FzE4 protein protein human Homo sapiens Human MAWRGAGPSVPGAPGGVGLSLGLLLQLLLLLGPARGFGDEEERRCDPIRISMCQNLGYNVTKMPNLVGHELQTDAELQLTTFTPLIQYGCSSQLQFFLCSVYVPMCTEKINIPIGPCGGMCLSVKRRCEPVLKEFGFAWPESLNCSKFPPQNDHNHMCMEGPGDEEVPLPHKTPIQPGEECHSVGTNSDQYIWVKRSLNCVLKCGYDAGLYSRSAKEFTDIWMAVWASLCFISTAFTVLTFLIDSSRFSYPERPIIFLSMCYNIYSIAYIVRLTVGRERISCDFEEAAEPVLIQEGLKNTGCAIIFLLMYFFGMASSIWWVILTLTWFLAAGLKWGHEAIEMHSSYFHIAAWAIPAVKTIVILIMRLVDADELTGLCYVGNQNLDALTGFVVAPLFTYLVIGTLFIAAGLVALFKIRSNLQKDGTKTDKLERLMVKIGVFSVLYTVPATCVIACYFYEISNWALFRYSADDSNMAVEMLKIFMSLLVGITSGMWIWSAKTLHTWQKCSNRLVNSGKVKREKRGNGWVKPGKGSETVV E0A83ECEC560A381 fzd1_rat Frizzled-1 Fzd1 protein protein rat Rattus norvegicus (Rat) Rat MAEEAVPSESRAAGRPSLELCAVALPGRREEVGHQDTAGHRRPRAHSRCWARGLLLLLWLLEAPLLLGVRAQPAGQVSGPGQQRPPPPQPQQGGQQYNGERGISIPDHGYCQPISIPLCTDIAYNQTIMPNLLGHTNQEDAGLEVHQFYPLVKVQCSAELKFFLCSMYAPVCTVLEQALPPCRSLCERAQGCEALMNKFGFQWPDTLKCEKFPVHGAGELCVGQNTSDKGTPTPSLLPEFWTSNPQHGGGGYRGGYPGGAGPVERGKFSCPRALRVPSYLNYHFLGEKDCGAPCEPTKVYGLMYFGPEELRFSRTWIGIWSVLCCASTLFTVLTYLVDMRRFSYPERPIIFLSGCYTAVAVAYIAGFLLEDRVVCNDKFAEDGARTVAQGTKKEGCTILFMMLYFFSMASSIWWVILSLTWFLAAGMKWGHEAIEANSQYFHLAAWAVPAIKTITILALGQVDGDVLSGVCFVGLNNVDALRGFVLAPLFVYLFIGTSFLLAGFVSLFRIRTIMKHDGTKTEKLEKLMVRIGVFSVLYTVPATIVIACYFYEQAFRDQWERSWVAQSCKSYAIPCPHLQGGGGVPPHPPMSPDFTVFMIKYLMTLIVGITSGFWIWSGKTLNSWRKFYTRLTNSKQGETTV D82E2C113E81B8B6 fzd1-fzd1-4 1 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait nucl lipotransfect nucleic acid transfection with liposome nucl lipotransfect region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1B, S1a,b, S2 IP in the presence of BS, a cell-impermeable cros-linker showed that the interaction takes place in the plasma membrane. fzd1-fzd2 1 4 unspecified role unspecified role prey prey myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role bait bait nucl lipotransfect nucleic acid transfection with liposome nucl lipotransfect region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1B, S1a,b IP in the presence of BS, a cell-impermeable cross linker showed that the interaction takes place at the plasma membrane. fzd1-fzd1-5 2 6 unspecified role unspecified role unspecified role unspecified role laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range certain certain sequence position certain renilla luciferase firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined This mutant lacks CRD (cysteine-rich domain): PMID: 11452312 6 unspecified role unspecified role unspecified role unspecified role laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1e, 2b I didn't annotate the Delta 360-381 sufficient to bind region of rat Frizzled 1. a), because I am not sure it is absolutely required and b) because according to Uniprot the C-terminal region containing the PDZ domain is 619-641, much different than 360-381stated in the paper (Fig. 2 legend) and 5th line from the bottom, left column page 54. Species is definetely rat. fzd1-fzd2-6 2 4 unspecified role unspecified role unspecified role unspecified role laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined renilla luciferase firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role unspecified role unspecified role laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1e fzd2-fzd2-4 2 4 unspecified role unspecified role unspecified role unspecified role laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role unspecified role unspecified role luciferase firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1e fzd1-fzd2-9 3 4 unspecified role unspecified role unspecified role unspecified role luciferase firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role unspecified role unspecified role gfp 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 proximity proximity close-contact colocalization neighbourhood interaction Fig. 1f fzd2-fzd2-6 3 4 unspecified role unspecified role unspecified role unspecified role renilla luciferase firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role unspecified role unspecified role gfp 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 proximity proximity close-contact colocalization neighbourhood interaction Fig. 1f fzd1-fzd1-7 3 6 unspecified role unspecified role unspecified role unspecified role gfp 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 unspecified role unspecified role renilla luciferase firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction Fig. 1f fzd1-fzd2-10 3 4 unspecified role unspecified role unspecified role unspecified role gfp 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 unspecified role unspecified role renilla luciferase firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction Fig. 1f fzd1-fzd1-6 2 6 unspecified role unspecified role unspecified role unspecified role laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role unspecified role unspecified role laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. S4c (labelled wrongly S4a in the main text) fzd2-fzd2-5 2 4 unspecified role unspecified role unspecified role unspecified role laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role unspecified role unspecified role laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. S4c (labelled wrongly S4a in the main text) fzd1-fzd2-7 2 4 unspecified role unspecified role unspecified role unspecified role laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role unspecified role unspecified role laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. S4c (labelled wrongly S4a in the main text) fzd1-fzd2-8 2 4 unspecified role unspecified role unspecified role unspecified role laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role unspecified role unspecified role laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. S4c (labelled wrongly S4a in the main text) fzd4-fzd4 2 5 unspecified role unspecified role unspecified role unspecified role nucl lipotransfect nucleic acid transfection with liposome nucl lipotransfect laczω fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region firefly luciferase protein tag firefly luciferase protein tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 unspecified role unspecified role unspecified role unspecified role nucl lipotransfect nucleic acid transfection with liposome nucl lipotransfect laczalpha fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 4a