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/ Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Cell metabolism 2010 imex curation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down tag visualisation tag visualisation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". Cholesterol Dissociation Assay:Cholesterol:NPC2-His10 was diluted 9-fold with buffer containing unlabelled cholesterol. After incubation the sample was applied to Ni-Agarose beads, centifuged and the supernatant assyed for radioactivity. mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu Accepted 2017-MAR-06 AT 08:35 GMT AT 08:35 GMT by ORCHARD Cell metabolism 2010 imex curation Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Cell metabolism 2010 imex curation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down tag visualisation tag visualisation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". Cholesterol binding Assay:Medium containing His tagged NPC2 or NCP1 was incubated with [3H]cholesterol, and the NPC-bound sterol was isolated by nickel chromatography. mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu Accepted 2017-MAR-06 AT 08:35 GMT AT 08:35 GMT by ORCHARD Cell metabolism 2010 imex curation Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Cell metabolism 2010 imex curation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down tag visualisation tag visualisation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". Cholesterol Transfer Assays:- Type "A": Assay contained [3H] lipid pre-bound to a donor protein. Acceptor protein is His-tagged and subsequently isolated by Ni-resin pulldown. Protein-bound [3H]lipid was eluted and quantified by scintillation counting. mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu Accepted 2017-MAR-06 AT 08:35 GMT AT 08:35 GMT by ORCHARD Cell metabolism 2010 imex curation Altered to Bait/Prey.; NCP2 "purified" removed for Exp. Prep. Donor/acceptors removed.; altered;cholesterol is the Prey as it is captured by the Bait.; altered:cholesterol is the Prey again. Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Cell metabolism 2010 imex curation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro liposome binding assay liposome binding assay tag visualisation tag visualisation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". Liposome Transfer Assay:Radiolabelled NPC1(NTD) was incubated with liposomes made from phosphatidylcholine and incorporating Texas Red dye. Components were separated on a Nickel-agarose column and amount of cholesterol transferred to liposomes was quantified by scintillation counting. The red dye shows the location of the liposomes. "PC" was prepared from egg yolk L-alpha--phosphatidyl-choline; it has fatty acid contents of approximately 33% 16:0 (palmitic), 13% 18:0 (stearic), 31% 18:1(oleic), and 15% 18:2 (linoleic) (other fatty acids being minor contributors). mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu Accepted 2017-MAR-06 AT 08:35 GMT AT 08:35 GMT by ORCHARD Cell metabolism 2010 imex curation Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Identification of surface residues on Niemann-Pick C2 essential for hydrophobic handoff of cholesterol to NPC1 in lysosomes. Cell metabolism 2010 imex curation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu 18-Apr-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip tag visualisation tag visualisation Wang ML., Motamed M., Infante RE., Abi-Mosleh L., Kwon HJ., Brown MS., Goldstein JL. Kwon-2009 publication pmid:19563754 -already in Intact, has some of the same assays as used here. I have assumed the NCP proteins have been purified - so interactions are often "Direct" - I have left ones with 3 interactors - +chol.) present as "physical". Cholesterol Transfer Assays Type "B": [3H]- cholesterol is pre-bound to a donor protein (NCP2) : it is transferred to acceptor NPC1(NTD)-LVPRGS-His8-FLAG. Anti-FLAG M2-agarose beads were used to isolate NCP1, and associated radioactivity counted. mike.brown@utsouthwestern.edu,joe.goldstein@utsouthwestern.edu Accepted 2017-MAR-06 AT 08:35 GMT AT 08:35 GMT by ORCHARD Cell metabolism 2010 imex curation phosphatidylcholine phosphatidylcholine 1,2-diacyl-glycero-3-phosphocholine a phosphatidylcholine (0) 2,3-bis(acyloxy)propyl 2-(trimethylammonio)ethyl phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cholesterol cholesterol small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) npc2_human NPC intracellular cholesterol transporter 2 NPC2 Niemann-Pick disease type C2 protein HE1 Human epididymis-specific protein 1 Epididymal secretory protein E1 protein protein human Homo sapiens Human MRFLAATFLLLALSTAAQAEPVQFKDCGSVDGVIKEVNVSPCPTQPCQLSKGQSYSVNVTFTSNIQSKSSKAVVHGILMGVPVPFPIPEPDGCKSGINCPIQKDKTYSYLNKLPVKSEYPSIKLVVEWQLQDDKNQSLFCWEIPVQIVSHL B141B611805DC910 npc1_human NPC intracellular cholesterol transporter 1 NPC1 Niemann-Pick C1 protein protein protein human Homo sapiens Human MTARGLALGLLLLLLCPAQVFSQSCVWYGECGIAYGDKRYNCEYSGPPKPLPKDGYDLVQELCPGFFFGNVSLCCDVRQLQTLKDNLQLPLQFLSRCPSCFYNLLNLFCELTCSPRQSQFLNVTATEDYVDPVTNQTKTNVKELQYYVGQSFANAMYNACRDVEAPSSNDKALGLLCGKDADACNATNWIEYMFNKDNGQAPFTITPVFSDFPVHGMEPMNNATKGCDESVDEVTAPCSCQDCSIVCGPKPQPPPPPAPWTILGLDAMYVIMWITYMAFLLVFFGAFFAVWCYRKRYFVSEYTPIDSNIAFSVNASDKGEASCCDPVSAAFEGCLRRLFTRWGSFCVRNPGCVIFFSLVFITACSSGLVFVRVTTNPVDLWSAPSSQARLEKEYFDQHFGPFFRTEQLIIRAPLTDKHIYQPYPSGADVPFGPPLDIQILHQVLDLQIAIENITASYDNETVTLQDICLAPLSPYNTNCTILSVLNYFQNSHSVLDHKKGDDFFVYADYHTHFLYCVRAPASLNDTSLLHDPCLGTFGGPVFPWLVLGGYDDQNYNNATALVITFPVNNYYNDTEKLQRAQAWEKEFINFVKNYKNPNLTISFTAERSIEDELNRESDSDVFTVVISYAIMFLYISLALGHMKSCRRLLVDSKVSLGIAGILIVLSSVACSLGVFSYIGLPLTLIVIEVIPFLVLAVGVDNIFILVQAYQRDERLQGETLDQQLGRVLGEVAPSMFLSSFSETVAFFLGALSVMPAVHTFSLFAGLAVFIDFLLQITCFVSLLGLDIKRQEKNRLDIFCCVRGAEDGTSVQASESCLFRFFKNSYSPLLLKDWMRPIVIAIFVGVLSFSIAVLNKVDIGLDQSLSMPDDSYMVDYFKSISQYLHAGPPVYFVLEEGHDYTSSKGQNMVCGGMGCNNDSLVQQIFNAAQLDNYTRIGFAPSSWIDDYFDWVKPQSSCCRVDNITDQFCNASVVDPACVRCRPLTPEGKQRPQGGDFMRFLPMFLSDNPNPKCGKGGHAAYSSAVNILLGHGTRVGATYFMTYHTVLQTSADFIDALKKARLIASNVTETMGINGSAYRVFPYSVFYVFYEQYLTIIDDTIFNLGVSLGAIFLVTMVLLGCELWSAVIMCATIAMVLVNMFGVMWLWGISLNAVSLVNLVMSCGISVEFCSHITRAFTVSMKGSRVERAEEALAHMGSSVFSGITLTKFGGIVVLAFAKSQIFQIFYFRMYLAMVLLGATHGLIFLPVLLSYIGPSVNKAKSCATEERYKGTERERLLNF DIP protein O15118 original sequence version: 157 DA1523E09822E5C7 npc2-cholesterol-1 2 7 unspecified role unspecified role prey prey region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait purified purified P61916:p.Val81Ala 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 V A Wed May 02 16:27:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81ala' Tue Jun 30 16:05:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Ala' P61916:p.Pro114Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain P A Wed May 02 15:09:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro114ala' Tue Jun 30 18:58:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro114Ala' P61916:p.Lys116Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain K A Wed May 02 15:56:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys116ala' Tue Jun 30 16:51:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys116Ala' P61916:p.Tyr119Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y A Wed May 02 19:29:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr119ala' Tue Jun 30 19:47:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr119Ala' P61916:p.Ile122Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain I A Wed May 02 17:42:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile122ala' Tue Jun 30 17:48:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile122Ala' P61916:p.Leu124Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain L A Wed May 02 18:57:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu124ala' Tue Jun 30 20:02:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu124Ala' P61916:p.Val81Asp 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 V D Wed May 02 16:31:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81asp' Tue Jun 30 17:10:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Asp' P61916:p.Gln45_Pro46delinsAlaAla mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain QP AA Wed May 02 19:45:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln_pro45-46ala_ala' Tue Jun 30 18:16:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln45_Pro46delinsAlaAla' P61916:p.Gly76Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain G A Wed May 02 15:27:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly76ala' Tue Jun 30 18:32:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly76Ala' P61916:p.Leu78Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain L A Wed May 02 18:39:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu78ala' Tue Jun 30 18:04:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu78Ala' P61916:p.[His75Ala;Ile77Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain H A certain certain sequence position certain certain certain sequence position certain I A Wed May 02 20:10:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his75ala,ile77ala' Tue Jun 30 17:26:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[His75Ala;Ile77Ala]' P61916:p.Phe85Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain F A Wed May 02 15:41:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe85ala' Tue Jun 30 18:21:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe85Ala' P61916:p.Ile87Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain I A Wed May 02 16:11:10 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile87ala' Tue Jun 30 16:11:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile87Ala' P61916:p.Pro88Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain P A Wed May 02 19:18:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro88ala' Tue Jun 30 17:10:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro88Ala' P61916:p.Pro100Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain P A Wed May 02 14:56:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'pro100ala' Tue Jun 30 18:19:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Pro100Ala' c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 direct interaction direct interaction 1 A/C, 2 B/C Mutations reducing binding >85% have been entered as shown by the blue bands in Fig 1C. npc1-npc2-1 4 9 unspecified role unspecified role neutral component neutral component purified purified ntd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component whole_molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 stimulator stimulator neutral component neutral component P61916:p.Met79Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain M A Wed May 02 19:12:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'met79ala' Tue Jun 30 20:24:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Met79Ala' P61916:p.Val81Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V A Wed May 02 15:19:47 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81ala' Tue Jun 30 19:09:52 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Ala' P61916:p.Val83Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain V A Wed May 02 19:01:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val83ala' Tue Jun 30 19:10:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val83Ala' crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 32 37 physical association physical association 1 B/D I have used CHEBI:64482 for PC as any more specific identifiers (e.g. CHEBI:57643) have associated +1 or _1 charges - and that is not mentioned for this mol, so have used higher term in ontology. Mutants showing transfer activity less than 40% of WT have been entered. It is not stated if NCP2 is tagged or not, in Figure 1 Liposome Transfer Assays. cholesterol-npc1-1 4 9 unspecified role unspecified role neutral component neutral component purified purified ntd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component whole_molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 stimulator stimulator neutral component neutral component purified purified P61916:p.Val81Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain V A Wed May 02 15:05:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81ala' Tue Jun 30 15:39:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Ala' P61916:p.Val81Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V D Wed May 02 16:35:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81asp' Tue Jun 30 18:38:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Asp' c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 50 45 physical association physical association 2 A I have used CHEBI:64482 for PC as any more specific identifiers (e.g. CHEBI:57643) have associated +1 or _1 charges - and that is not mentioned for this mol, so have used higher term in ontology. npc2-cholesterol-2 1 7 unspecified role unspecified role prey prey whole_molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait purified purified P61916:p.Val81Asp 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 V D Wed May 02 15:06:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81asp' Tue Jun 30 16:40:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Asp' c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 63 58 physical association physical association 2D cholesterol-npc1-2 5 8 unspecified role unspecified role unspecified role unspecified role purified purified c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P61916:p.Val81Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain V A Wed May 02 20:13:08 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81ala' Tue Jun 30 18:48:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Ala' P61916:p.Val81Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V D Wed May 02 17:11:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81asp' Tue Jun 30 15:36:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Asp' whole_molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 9 unspecified role unspecified role bait bait purified purified ntd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 7 unspecified role unspecified role prey prey whole_molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 75 69 physical association physical association 2E npc1-cholesterol-3 2 7 unspecified role unspecified role prey prey region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait purified purified ntd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range O15118:p.Leu175_Leu176delinsGlnGln 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 LL QQ Wed May 02 20:29:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_leu175-176gln_gln' Tue Jun 30 19:30:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu175_Leu176delinsGlnGln' direct interaction direct interaction 4A cholesterol-npc1-3 3 8 unspecified role unspecified role unspecified role unspecified role purified purified c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range P61916:p.Val81Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain V A Wed May 02 19:31:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81ala' Tue Jun 30 18:18:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Ala' P61916:p.Val81Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain V D Wed May 02 16:51:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val81asp' Tue Jun 30 18:46:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val81Asp' whole_molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 9 unspecified role unspecified role bait bait purified purified ntd sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range O15118:p.Leu175_Leu176delinsGlnGln mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain LL QQ Wed May 02 16:30:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu_leu175-176gln_gln' Tue Jun 30 16:00:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu175_Leu176delinsGlnGln' crigr-cho_k1 Cricetulus griseus ovary K1 cho_k1 Chinese Hamster Ovary K1 7 unspecified role unspecified role prey prey whole_molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 3h radiolabel 3h radiolabel 3H tritium H3 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 90 97 physical association physical association 4B