Molecular Connections Molecular Connections Pvt. Ltd. 2/2, Kariappa Road, Basavanagudi, Bangalore -560 004 www.molecularconnections.com www.molecularconnections.com gaugel-2012-2 Defining regulatory and phosphoinositide-binding sites in the human WIPI-1 beta-propeller responsible for autophagosomal membrane localization downstream of mTORC1 inhibition. in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay anti tag western anti tag western blot anti tag western Gaugel A., Bakula D., Hoffmann A., Proikas-Cezanne T. tassula.proikas-cezanne@uni-tuebingen.de J Mol Signal 2012 imex curation Molecular Connections Pvt. Ltd. Only protein-protein interactions gaugel-2012-1 Defining regulatory and phosphoinositide-binding sites in the human WIPI-1 beta-propeller responsible for autophagosomal membrane localization downstream of mTORC1 inhibition. human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference confocal microscopy confocal microscopy immunostaining immunostaining Gaugel A., Bakula D., Hoffmann A., Proikas-Cezanne T. tassula.proikas-cezanne@uni-tuebingen.de J Mol Signal 2012 imex curation Molecular Connections Pvt. Ltd. Only protein-protein interactions pi3p 1-phosphatidyl-1D-myo-inositol 3-phosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) wipi1_human WD repeat domain phosphoinositide-interacting protein 1 WD40 repeat protein interacting with phosphoinositides of 49 kDa Atg18 protein homolog WIPI1 WIPI49 protein protein human Homo sapiens Human MEAEAADAPPGGVESALSCFSFNQDCTSLATGTKAGYKLFSLSSVEQLDQVHGSNEIPDVYIVERLFSSSLVVVVSHTKPRQMNVYHFKKGTEICNYSYSSNILSIRLNRQRLLVCLEESIYIHNIKDMKLLKTLLDIPANPTGLCALSINHSNSYLAYPGSLTSGEIVLYDGNSLKTVCTIAAHEGTLAAITFNASGSKLASASEKGTVIRVFSVPDGQKLYEFRRGMKRYVTISSLVFSMDSQFLCASSNTETVHIFKLEQVTNSRPEEPSTWSGYMGKMFMAATNYLPTQVSDMMHQDRAFATARLNFSGQRNICTLSTIQKLPRLLVASSSGHLYMYNLDPQDGGECVLIKTHSLLGSGTTEENKENDLRPSLPQSYAATVARPSASSASTVPGYSEDGGALRGEVIPEHEFATGPVCLDDENEFPPIILCRGNQKGKTKQS 1-phosphatidyl-1d-myo-inositol 3,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 3,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) atg12_human Ubiquitin-like protein ATG12 Autophagy-related protein 12 ATG12 APG12 APG12L protein protein human Homo sapiens Human MAEEPQSVLQLPTSIAAGGEGLTDVSPETTTPEPPSSAAVSPGTEEPAGDTKKKIDILLKAVGDTPIMKTKKWAVERTRTIQGLIDFIKKFLKLVASEQLFIYVNQSFAPSPDQEVGTLYECFGSDGKLVLHYCKSQAWG 1_phosphatidyl_1d_myo_inositol_3_phosphate _wipi1 1328978 n/a 1328980 unspecified role unspecified role bait bait n/a WIPI-1 1328981 unspecified role unspecified role prey prey Q5MNZ9:p.Arg112Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false n-term 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 false GFP construct details obtained from back reference- PMID-15602573 Q5MNZ9:p.Arg226_Arg227delinsAlaAla mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.His185Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Ser203Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Ser205Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Gly208Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Thr209Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Arg212Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Arg226Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Arg227Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Gly228Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Ser251Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.Thr255Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q5MNZ9:p.His257Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference physical association physical association false false false figs. 6, 7 U2OS cells transiently express GFP-WIPI-1 or GFP-tagged mutant WIPI-1 variants the following mutations on WIPI1 did not nullify the ability to localize at autophagosomal membranes-Asn23, Gln24, Asp25, Glu64, Arg107, Arg110, Arg112, Gly198, Glu224, Phe225, Ser250, Ser335, Gly336 1_phosphatidyl_1d_myo_inositol_3_5_bisphosphate-wipi1 1328978 n/a 1328982 unspecified role unspecified role bait bait n/a WIPI-1 1328981 unspecified role unspecified role prey prey n-term 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 false GFP construct details obtained from back reference- PMID-15602573 Q5MNZ9:p.Arg226_Arg227delinsAlaAla mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference physical association physical association false false false figs. 6, 8 U2OS cells transiently express GFP-WIPI-1 or GFP-tagged mutant WIPI-1 variants the following mutations on WIPI1 did not nullify the ability to localize at autophagosomal membranes-Asn23, Gln24, Asp25, Glu64, Arg107, Arg110, Arg112, Gly198, Glu224, Phe225, Ser250, Ser335, Gly336 atg12-wipi1-1 1328979 n/a WIPI-1 1328981 unspecified role unspecified role neutral component neutral component n-term 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 false GFP construct details obtained from back reference- PMID-15602573 human-u2-os Human U2-OS osteosarcoma cell line u2-os U2-OS osteocaroma cell line Not in CAPRI and no primary literature reference n/a 1328983 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false fig. 4 U2OS cells transiently express GFP-WIPI-1 or GFP-tagged mutant WIPI-1 variants the following mutations on WIPI1 did not nullify the ability to localize at autophagosomal membranes-Asn23, Gln24, Asp25, Glu64, Arg107, Arg110, Arg112, Gly198, Glu224, Phe225, Ser250, Ser335, Gly336