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/ ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. Nature (0028-0836) 2006 imex curation Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-purkinje cell Mus musculus brain-purkinje_cell Purkinje cell Purkinje cells can be found either as 1] Purkinje fibers in the inner ventricular walls of the heart, that conduct an electrical stimulus that enables the heart to contract in a coordinated fashion - or 2]Purkinje neurons, a class of GABAergic neurons located in the cerebellar cortex. fluorescence imaging fluorescence microscopy fluorescence imaging antibody detection identification by antibody antibody detection Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de Accepted 2006-AUG-25 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nature (0028-0836) 2006 Only protein-protein interactions imex curation ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. Nature (0028-0836) 2006 imex curation Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-osteoclast Mouse osteoclast osteoclast Osteoclast fluorescence imaging fluorescence microscopy fluorescence imaging antibody detection identification by antibody antibody detection Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de Accepted 2006-AUG-25 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nature (0028-0836) 2006 Only protein-protein interactions imex curation ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. Nature (0028-0836) 2006 imex curation Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de 24-JAN-2007: Contacted by JYOTI. 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. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de Accepted 2006-AUG-25 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nature (0028-0836) 2006 Only protein-protein interactions imex curation ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. Nature (0028-0836) 2006 imex curation Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-fibroblast Mus musculus fibroblast Fibroblast fluorescence imaging fluorescence microscopy fluorescence imaging antibody detection identification by antibody antibody detection Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de Accepted 2006-AUG-25 by DAVET. Nature (0028-0836) 2006 Only protein-protein interactions imex curation ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. ClC-7 requires Ostm1 as a beta-subunit to support bone resorption and lysosomal function. Nature (0028-0836) 2006 imex curation Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-brain Mus musculus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lange PF., Wartosch L., Jentsch TJ., Fuhrmann JC. BioCreative - Critical Assessment of Information Extraction systems in Biology Jentsch@zmnh.uni-hamburg.de Accepted 2006-AUG-25 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nature (0028-0836) 2006 Only protein-protein interactions imex curation lamp1_mouse Lysosome-associated membrane glycoprotein 1 Lysosomal membrane glycoprotein A 120 kDa lysosomal membrane glycoprotein LGP-120 P2B CD107 antigen-like family member A Lamp1 Lamp-1 protein protein mouse Mus musculus Mouse MAAPGARRPLLLLLLAGLAHGASALFEVKNNGTTCIMASFSASFLTTYETANGSQIVNISLPASAEVLKNGSSCGKENVSDPSLTITFGRGYLLTLNFTKNTTRYSVQHMYFTYNLSDTEHFPNAISKEIYTMDSTTDIKADINKAYRCVSDIRVYMKNVTVVLRDATIQAYLSSGNFSKEETHCTQDGPSPTTGPPSPSPPLVPTNPTVSKYNVTGNNGTCLLASMALQLNITYLKKDNKTVTRAFNISPNDTSSGSCGINLVTLKVENKNRALELQFGMNASSSLFFLQGVRLNMTLPDALVPTFSISNHSLKALQATVGNSYKCNTEEHIFVSKMLSLNVFSVQVQAFKVDSDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKRSHAGYQTI C1BD373548BB9655 clcn7_mouse H(+)/Cl(-) exchange transporter 7 Clcn7 Clc7 Chloride channel protein 7 Chloride channel 7 alpha subunit protein protein mouse Mus musculus Mouse MANVSKKVSWSGRDRDDEEGAPLLRRTGQPDEETPLLNGAGPGARQSHSALFRIGQMNNVELDDELLDPEVDPPHTFPKEIPHNEKLLSLKYESLDYDNSENQLFLEEERRINHTAFRTVEIKRWVICALIGILTGLVACFIDIVVENLAGLKYRVIKDNIDKFTEKGGLSFSLLLWATLNSAFVLVGSVIVAFIEPVAAGSGIPQIKCFLNGVKIPHVVRLKTLVIKVSGVILSVVGGLAVGKEGPMIHSGSVIAAGISQGRSTSLKRDFKIFEYFRRDTEKRDFVSAGAAAGVSAAFGAPVGGVLFSLEEGASFWNQFLTWRIFFASMISTFTLNFVLSIYHGNMWDLSSPGLINFGRFDSEKMAYTIHEIPVFIAMGVVGGILGAVFNALNYWLTMFRIRYIHRPCLQVIEAMLVAAVTATVAFVLIYSSRDCQPLQGSSMSYPLQLFCADGEYNSMAAAFFNTPEKSVVSLFHDPPGSYNPMTLGLFTLVYFFLACWTYGLTVSAGVFIPSLLIGAAWGRLFGISLSYLTGAAIWADPGKYALMGAAAQLGGIVRMTLSLTVIMMEATSNVTYGFPIMLVLMTAKIVGDVFIEGLYDMHIQLQSVPFLHWEAPVTSHSLTAREVMSTPVTCLRRREKVGIIVDVLSDTASNHNGFPVVEDVGDTQPARLQGLILRSQLIVLLKHKVFVERSNMGLVQRRLRLKDFRDAYPRFPPIQSIHVSQDERECTMDLSEFMNPSPYTVPQEASLPRVFKLFRALGLRHLVVVDNHNQVVGLVTRKDLARYRLGKGGLEELSLAQT A7D6DA5791DAA48C ostm1_mouse Osteopetrosis-associated transmembrane protein 1 Grey-lethal protein Ostm1 Gl Chloride channel 7 beta subunit protein protein mouse Mus musculus Mouse MARDAELARSSGWPWRWLPALLLLQLLRWRCALCALPFTSSRHPGFADLLSEQQLLEVQDLTLSLLQGGGLGPLSLLPPDLPDLEPECRELLMDFANSSAELTACMVRSARPVRLCQTCYPLFQQVAIKMDNISRNIGNTSEGPRCGGSLLTADRMQIVLMVSEFFNSTWQEANCANCLTNNGEDLSNNTEDFLSLFNKTLACFEHNLQGHTYSLLPPKNYSEVCRNCKEAYKNLSLLYSQMQKLNGLENKAEPETHLCIDVEDAMNITRKLWSRTFNCSVTCSDTVSVVAVSVFILFLPVVFYLSSFLHSEQKKRKLILPKRLKSSTSFANIQENAT 73EF8EE9ED760C08 ostm1-lamp1-1 4 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization In cultured primary fibroblasts, Ostm1 co-localized with Lamp-1, a marker for late endosomes and lysosomes (Fig. 2a). 1A ostm1-lamp1-2 4 6 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization Co-transfection with ClC-7 restored a punctate Ostm1 staining pattern that largely co-stained with Lamp-1 (Fig. 2c) and ClC-7 (Fig. 2d). Q8BGT0 transfection; Colocalisation required cotransfection with Clc-7 (O70496) 2C ostm1-clcn7-1 4 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization Co-transfection with ClC-7 restored a punctate Ostm1 staining pattern that largely co-stained with Lamp-1 (Fig. 2c) and ClC-7 (Fig. 2d). O70496 transfection; O70496 transfection; 2D ostm1-clcn7-2 5 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association ClC-7 was efficiently co-immunoprecipitated from the brain with Ostm1, and vice versa (Fig. 3d). 3D ostm1-clcn7-3 5 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association ClC-7 was efficiently co-immunoprecipitated from the brain with Ostm1, and vice versa (Fig. 3d). 3D Molecular weight analysis suggested a cleaved, lysosomal Ostm1 fragment was pulled down by the Clcn7 antibody. ostm1-clcn7-4 3 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association Co-immunoprecipitation performed with transfected cells in which only the large form of Ostm1 could be detected showed that this putative ER form also interacted with ClC-7 (Supplementary Fig. S2). S2 Molecular weight analysis suggested a higher molecular weight form of Ostm1 was pulled down by the Clcn7 antibody. Q8BGT0 transfection; ostm1-clcn7-5 1 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization a, b, Immunofluorescence of cerebellar Purkinje cells. a, Ostm1 co-localized with ClC-7 in late endosomes/lysosomes4 of wild-type cells. 4A ostm1-clcn7-6 2 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization Immunofluorescence of osteoclasts in situ. Ostm1 and ClC-7 co-localized in the ruffled border (c), as did ClC-7 and the a3 proton-pump subunit (d). 4C tcirg1-clcn7-1 2 8 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component colocalization colocalization Immunofluorescence of osteoclasts in situ. Ostm1 and ClC-7 co-localized in the ruffled border (c), as did ClC-7 and the a3 proton-pump subunit (d). 4D & 4E