UniProt UniProt http://www.uniprot.org barbelanne-2013-2 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 tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Barbelanne M., Song J., Ahmadzai M., Tsang WY. Rare diseases - Interactions investigated in the context of Rare genetic disease william.tsang@ircm.qc.ca Human molecular genetics 2013 Only protein-protein interactions imex curation imex curation barbelanne-2013-3 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 tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Barbelanne M., Song J., Ahmadzai M., Tsang WY. Rare diseases - Interactions investigated in the context of Rare genetic disease Interaction using Flag-CaM1234 as bait have been omitted as details of the mutations cannot be traced. william.tsang@ircm.qc.ca Human molecular genetics 2013 Only protein-protein interactions imex curation imex curation barbelanne-2013-1 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 Barbelanne M., Song J., Ahmadzai M., Tsang WY. Rare diseases - Interactions investigated in the context of Rare genetic disease william.tsang@ircm.qc.ca Human molecular genetics 2013 Only protein-protein interactions imex curation imex curation barbelanne-2013-7 human-htert_rpe1 Homo sapiens htert_rpe1 htert_rpe1 fluorescence imaging fluorescence microscopy fluorescence imaging immunocytochemistry immunocytochemistry Barbelanne M., Song J., Ahmadzai M., Tsang WY. Rare diseases - Interactions investigated in the context of Rare genetic disease william.tsang@ircm.qc.ca Human molecular genetics 2013 Only protein-protein interactions imex curation imex curation barbelanne-2013-4 in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting anti tag western anti tag western blot anti tag western Barbelanne M., Song J., Ahmadzai M., Tsang WY. Rare diseases - Interactions investigated in the context of Rare genetic disease william.tsang@ircm.qc.ca Human molecular genetics 2013 Only protein-protein interactions imex curation imex curation barbelanne-2013-5 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 western blot western blot Immuno blot Immuno blot Barbelanne M., Song J., Ahmadzai M., Tsang WY. Rare diseases - Interactions investigated in the context of Rare genetic disease william.tsang@ircm.qc.ca Human molecular genetics 2013 Only protein-protein interactions imex curation imex curation barbelanne-2013-6 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Barbelanne M., Song J., Ahmadzai M., Tsang WY. Rare diseases - Interactions investigated in the context of Rare genetic disease william.tsang@ircm.qc.ca Human molecular genetics 2013 Only protein-protein interactions imex curation imex curation ce290_human Centrosomal protein of 290 kDa CEP290 KIAA0373 NPHP6 Nephrocystin-6 Tumor antigen se2-2 Cancer/testis antigen 87 BBS14 Bardet-Biedl syndrome 14 protein protein protein human Homo sapiens Human MPPNINWKEIMKVDPDDLPRQEELADNLLISLSKVEVNELKSEKQENVIHLFRITQSLMKMKAQEVELALEEVEKAGEEQAKFENQLKTKVMKLENELEMAQQSAGGRDTRFLRNEICQLEKQLEQKDRELEDMEKELEKEKKVNEQLALRNEEAENENSKLRRENKRLKKKNEQLCQDIIDYQKQIDSQKETLLSRRGEDSDYRSQLSKKNYELIQYLDEIQTLTEANEKIEVQNQEMRKNLEESVQEMEKMTDEYNRMKAIVHQTDNVIDQLKKENDHYQLQVQELTDLLKSKNEEDDPIMVAVNAKVEEWKLILSSKDDEIIEYQQMLHNLREKLKNAQLDADKSNVMALQQGIQERDSQIKMLTEQVEQYTKEMEKNTCIIEDLKNELQRNKGASTLSQQTHMKIQSTLDILKEKTKEAERTAELAEADAREKDKELVEALKRLKDYESGVYGLEDAVVEIKNCKNQIKIRDREIEILTKEINKLELKISDFLDENEALRERVGLEPKTMIDLTEFRNSKHLKQQQYRAENQILLKEIESLEEERLDLKKKIRQMAQERGKRSATSGLTTEDLNLTENISQGDRISERKLDLLSLKNMSEAQSKNEFLSRELIEKERDLERSRTVIAKFQNKLKELVEENKQLEEGMKEILQAIKEMQKDPDVKGGETSLIIPSLERLVNAIESKNAEGIFDASLHLKAQVDQLTGRNEELRQELRESRKEAINYSQQLAKANLKIDHLEKETSLLRQSEGSNVVFKGIDLPDGIAPSSASIINSQNEYLIHLLQELENKEKKLKNLEDSLEDYNRKFAVIRHQQSLLYKEYLSEKETWKTESKTIKEEKRKLEDQVQQDAIKVKEYNNLLNALQMDSDEMKKILAENSRKITVLQVNEKSLIRQYTTLVELERQLRKENEKQKNELLSMEAEVCEKIGCLQRFKEMAIFKIAALQKVVDNSVSLSELELANKQYNELTAKYRDILQKDNMLVQRTSNLEHLECENISLKEQVESINKELEITKEKLHTIEQAWEQETKLGNESSMDKAKKSITNSDIVSISKKITMLEMKELNERQRAEHCQKMYEHLRTSLKQMEERNFELETKFAELTKINLDAQKVEQMLRDELADSVSKAVSDADRQRILELEKNEMELKVEVSKLREISDIARRQVEILNAQQQSRDKEVESLRMQLLDYQAQSDEKSLIAKLHQHNVSLQLSEATALGKLESITSKLQKMEAYNLRLEQKLDEKEQALYYARLEGRNRAKHLRQTIQSLRRQFSGALPLAQQEKFSKTMIQLQNDKLKIMQEMKNSQQEHRNMENKTLEMELKLKGLEELISTLKDTKGAQKVINWHMKIEELRLQELKLNRELVKDKEEIKYLNNIISEYERTISSLEEEIVQQNKFHEERQMAWDQREVDLERQLDIFDRQQNEILNAAQKFEEATGSIPDPSLPLPNQLEIALRKIKENIRIILETRATCKSLEEKLKEKESALRLAEQNILSRDKVINELRLRLPATAEREKLIAELGRKEMEPKSHHTLKIAHQTIANMQARLNQKEEVLKKYQRLLEKAREEQREIVKKHEEDLHILHHRLELQADSSLNKFKQTAWDLMKQSPTPVPTNKHFIRLAEMEQTVAEQDDSLSSLLVKLKKVSQDLERQREITELKVKEFENIKLQLQENHEDEVKKVKAEVEDLKYLLDQSQKESQCLKSELQAQKEANSRAPTTTMRNLVERLKSQLALKEKQQKALSRALLELRAEMTAAAEERIISATSQKEAHLNVQQIVDRHTRELKTQVEDLNENLLKLKEALKTSKNRENSLTDNLNDLNNELQKKQKAYNKILREKEEIDQENDELKRQIKRLTSGLQGKPLTDNKQSLIEELQRKVKKLENQLEGKVEEVDLKPMKEKNAKEELIRWEEGKKWQAKIEGIRNKLKEKEGEVFTLTKQLNTLKDLFAKADKEKLTLQRKLKTTGMTVDQVLGIRALESEKELEELKKRNLDLENDILYMRAHQALPRDSVVEDLHLQNRYLQEKLHALEKQFSKDTYSKPSISGIESDDHCQREQELQKENLKLSSENIELKFQLEQANKDLPRLKNQVRDLKEMCEFLKKEKAEVQRKLGHVRGSGRSGKTIPELEKTIGLMKKVVEKVQRENEQLKKASGILTSEKMANIEQENEKLKAELEKLKAHLGHQLSMHYESKTKGTEKIIAENERLRKELKKETDAAEKLRIAKNNLEILNEKMTVQLEETGKRLQFAESRGPQLEGADSKSWKSIVVTRMYETKLKELETDIAKKNQSITDLKQLVKEATEREQKVNKYNEDLEQQIKILKHVPEGAETEQGLKRELQVLRLANHQLDKEKAELIHQIEANKDQSGAESTIPDADQLKEKIKDLETQLKMSDLEKQHLKEEIKKLKKELENFDPSFFEEIEDLKYNYKEEVKKNILLEEKVKKLSEQLGVELTSPVAASEEFEDEEESPVNFPIY calm_human Calmodulin CALM1 CALM CAM CAM1 CALM2 CAMB CALM3 CAM3 CAMC CAMIII CAM2 CALML2 protein protein human Homo sapiens Human MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK iqcb1_human IQ calmodulin-binding motif-containing protein 1 KIAA0036 NPHP5 Nephrocystin-5 IQCB1 p53 and DNA damage-regulated IQ motif protein OK/SW-cl.85 protein protein human Homo sapiens Human MKPTGTDPRILSIAAEVAKSPEQNVPVILLKLKEIINITPLGSSELKKIKQDIYCYDLIQYCLLVLSQDYSRIQGGWTTISQLTQILSHCCVGLEPGEDAEEFYNELLPSAAENFLVLGRQLQTCFINAAKAEEKDELLHFFQIVTDSLFWLLGGHVELIQNVLQSDHFLHLLQADNVQIGSAVMMMLQNILQINSGDLLRIGRKALYSILDEVIFKLFSTPSPVIRSTATKLLLLMAESHQEILILLRQSTCYKGLRRLLSKQETGTEFSQELRQLVGLLSPMVYQEVEEQKLHQAACLIQAYWKGFQTRKRLKKLPSAVIALQRSFRSKRSKMLLEINRQKEEEDLKLQLQLQRQRAMRLSRELQLSMLEIVHPGQVEKHYREMEEKSALIIQKHWRGYRERKNFHQQRQSLIEYKAAVTLQRAALKFLAKCRKKKKLFAPWRGLQELTDARRVELKKRVDDYVRRHLGSPMSDVVSRELHAQAQERLQHYFMGRALEERAQQHREALIAQISTNVEQLMKAPSLKEAEGKEPELFLSRSRPVAAKAKQAHLTTLKHIQAPWWKKLGEESGDEIDVPKDELSIELENLFIGGTKPP calm_rat Calmodulin Calm1 Calm Cam Cam1 Calm2 Cam2 Camb Calm3 Cam3 Camc CaMII protein protein rat Rattus norvegicus (Rat) Rat MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK iqcb1-calm1-2 1937280 N/A 1937287 unspecified role unspecified role prey prey N/A 1937288 unspecified role unspecified role prey prey N/A 1937289 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q15051:p.Ala547Lys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15051:p.Lys550Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15051:p.Gln551Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15051:p.Lys548Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false association association false false false 3B iqcb1-calm1-3 1937280 N/A 1937288 unspecified role unspecified role prey prey N/A 1937289 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection Q15051:p.Ala547Lys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15051:p.Lys548Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15051:p.Ala549Lys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false no interaction with CEP290 region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q15051:p.Ile393Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false mutation retains interaction with CEP290 IQ regions 1-3 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false Single IQ motif deletion does not change interaction with calmodulin and CEP290. Triple deletion IQ123 disrupts the interaction with calmodulin but retains interaction with CEP290. Q15051:p.Lys550Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15051:p.Gln551Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 3B, 5A,6 Complete loss of binding was achieved only when all three IQ motifs were deleted. Two disease associated mutations of NPHP5 - F142fsX146 (424–425delTT,) and R332X result in proteins that did not interact with CAM. cep290-calm1-3 1937280 N/A 1937287 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false also binds IQCB1/NPHP5 N/A 1937288 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate association association false false false supplemental fig. S3A iqcb1-calm1-5 1937280 N/A 1937287 unspecified role unspecified role prey prey N/A 1937289 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false deletion disrupts interaction with CEP290 but retains interaction with calmodulin region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false deletion disrupts interaction with CEP290 but retains interaction with calmodulin Q15051:p.Ala549Lys mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false mutation disrupts interaction with CEP290 but retains interaction with calmodulin physical association physical association false false false 3Bi, 6 Several disease associated NPHP5 mutations -(F142fsX146, R332X, R461X, R502X and H506fsX518) result in proteins that do not bind to CEP290 cep290-iqcb1-2 1937281 N/A 1937287 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 1937289 unspecified role unspecified role prey prey nucl transfection nucleic acid transfection nucl transfection region 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 physical association physical association false false false supplemental fig. S3A cep290-calm1-2 1937281 N/A 1937287 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 1937288 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate N/A 1937289 unspecified role unspecified role prey prey nucl transfection nucleic acid transfection nucl transfection region 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 association association false false false supplemental fig. S3A calm1-cep290-1 1937281 N/A 1937287 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate N/A 1937290 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 1937289 unspecified role unspecified role prey prey nucl transfection nucleic acid transfection nucl transfection region 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 association association false false false supplemental fig. S5B calm1-cep290-3 1937281 N/A 1937287 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey cell lysate cell lysate N/A 1937290 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false supplemental fig. S5Bi iqcb1-calm1-1 1937282 N/A 1937287 unspecified role unspecified role prey prey N/A 1937288 unspecified role unspecified role prey prey N/A 1937289 unspecified role unspecified role bait bait association association false false false 3A cep290-calm1-1 1937282 N/A 1937287 unspecified role unspecified role bait bait N/A 1937288 unspecified role unspecified role prey prey N/A 1937289 unspecified role unspecified role prey prey association association false false false 3A 25345-1 1937283 N/A 1937287 unspecified role unspecified role neutral component neutral component N/A 1937289 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 1D cep290-iqcb1-5 1937283 N/A 1937287 unspecified role unspecified role neutral component neutral component N/A 1937289 unspecified role unspecified role neutral component neutral component region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Supp 5D cep290-iqcb1-3 1937284 N/A 1937287 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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. N/A 1937289 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false supplemental fig. S3c calm1-cep290-2 1937285 N/A 1937287 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey N/A 1937288 unspecified role unspecified role prey prey N/A 1937289 unspecified role unspecified role bait bait nucl transfection nucleic acid transfection nucl transfection region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false supplemental fig S5A EGTA iqcb1-calm1-4 1937286 N/A 1937288 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) N/A 1937289 unspecified role unspecified role bait bait purified purified 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false supplemental fig S5C