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/ Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. Proceedings of the National Academy of Sciences of the United States of America 2016 imex curation Li X., Saha P., Li J., Blobel G., Pfeffer SR. xli05@rockefeller.edu,blobel@rockefeller.edu,pfeffer@stanford.edu Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Li X., Saha P., Li J., Blobel G., Pfeffer SR. xli05@rockefeller.edu,blobel@rockefeller.edu,pfeffer@stanford.edu Accepted 2017-FEB-21 AT 12:21 GMT AT 12:21 GMT by ORCHARD Proceedings of the National Academy of Sciences of the United States of America 2016 imex curation Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. Clues to the mechanism of cholesterol transfer from the structure of NPC1 middle lumenal domain bound to NPC2. Proceedings of the National Academy of Sciences of the United States of America 2016 imex curation Li X., Saha P., Li J., Blobel G., Pfeffer SR. xli05@rockefeller.edu,blobel@rockefeller.edu,pfeffer@stanford.edu Only protein-protein interactions in vitro In vitro mst microscale thermophoresis mst predetermined predetermined participant predetermined Li X., Saha P., Li J., Blobel G., Pfeffer SR. xli05@rockefeller.edu,blobel@rockefeller.edu,pfeffer@stanford.edu Accepted 2017-FEB-21 AT 12:21 GMT AT 12:21 GMT by ORCHARD Proceedings of the National Academy of Sciences of the United States of America 2016 imex curation npc1_mouse NPC intracellular cholesterol transporter 1 Niemann-Pick C1 protein Npc1 protein protein mouse Mus musculus Mouse MGAHHPALGLLLLLLCPAQVFSQSCVWYGECGIATGDKRYNCKYSGPPKPLPKDGYDLVQELCPGLFFDNVSLCCDIQQLQTLKSNLQLPLQFLSRCPSCFYNLMTLFCELTCSPHQSQFLNVTATEDYFDPKTQENKTNVKELEYFVGQSFANAMYNACRDVEAPSSNEKALGLLCGRDARACNATNWIEYMFNKDNGQAPFTIIPVFSDLSILGMEPMRNATKGCNESVDEVTGPCSCQDCSIVCGPKPQPPPPPMPWRIWGLDAMYVIMWVTYVAFLFVFFGALLAVWCHRRRYFVSEYTPIDSNIAFSVNSSDKGEASCCDPLGAAFDDCLRRMFTKWGAFCVRNPTCIIFFSLAFITVCSSGLVFVQVTTNPVELWSAPHSQARLEKEYFDKHFGPFFRTEQLIIQAPNTSVHIYEPYPAGADVPFGPPLNKEILHQVLDLQIAIESITASYNNETVTLQDICVAPLSPYNKNCTIMSVLNYFQNSHAVLDSQVGDDFYIYADYHTHFLYCVRAPASLNDTSLLHGPCLGTFGGPVFPWLVLGGYDDQNYNNATALVITFPVNNYYNDTERLQRAWAWEKEFISFVKNYKNPNLTISFTAERSIEDELNRESNSDVFTVIISYVVMFLYISLALGHIQSCSRLLVDSKISLGIAGILIVLSSVACSLGIFSYMGMPLTLIVIEVIPFLVLAVGVDNIFILVQTYQRDERLQEETLDQQLGRILGEVAPTMFLSSFSETSAFFFGALSSMPAVHTFSLFAGMAVLIDFLLQITCFVSLLGLDIKRQEKNHLDILCCVRGADDGQGSHASESYLFRFFKNYFAPLLLKDWLRPIVVAVFVGVLSFSVAVVNKVDIGLDQSLSMPNDSYVIDYFKSLAQYLHSGPPVYFVLEEGYNYSSRKGQNMVCGGMGCDNDSLVQQIFNAAELDTYTRVGFAPSSWIDDYFDWVSPQSSCCRLYNVTHQFCNASVMDPTCVRCRPLTPEGKQRPQGKEFMKFLPMFLSDNPNPKCGKGGHAAYGSAVNIVGDDTYIGATYFMTYHTILKTSADYTDAMKKARLIASNITETMRSKGSDYRVFPYSVFYVFYEQYLTIIDDTIFNLSVSLGSIFLVTLVVLGCELWSAVIMCITIAMILVNMFGVMWLWGISLNAVSLVNLVMSCGISVEFCSHITRAFTMSTKGSRVSRAEEALAHMGSSVFSGITLTKFGGIVVLAFAKSQIFEIFYFRMYLAMVLLGATHGLIFLPVLLSYIGPSVNKAKRHTTYERYRGTERERLLNF 3B42230AAC86764E npc2_bovin NPC intracellular cholesterol transporter 2 Epididymal secretory protein E1 NPC2 Niemann Pick type C2 protein homolog EPV20 protein protein bovin Bos taurus (Bovine) Bovine MRFLTVAFLFLALSASALAEPVKFKDCGSWVGVIKEVNVSPCPTQPCKLHRGQSYSVNVTFTSNTQSQSSKAVVHGIVMGIPVPFPIPESDGCKSGIRCPIEKDKTYNYVNKLPVKNEYPSIKVVVEWELTDDKNQRFFCWQIPIEVEA 91156DC7E805B655 p61916-1 NPC intracellular cholesterol transporter 2 Human epididymis-specific protein 1 Niemann-Pick disease type C2 protein Epididymal secretory protein E1 NPC2 HE1 protein protein human Homo sapiens Human MRFLAATFLLLALSTAAQAEPVQFKDCGSVDGVIKEVNVSPCPTQPCQLSKGQSYSVNVTFTSNIQSKSSKAVVHGILMGVPVPFPIPEPDGCKSGINCPIQKDKTYSYLNKLPVKSEYPSIKLVVEWQLQDDKNQSLFCWEIPVQIVSHL B141B611805DC910 cholesterol sulfate cholesterol sulfate CHOLEST-5-EN-3-YL HYDROGEN SULFATE Cholest-5-en-3beta-ol sulfate cholesterol sulfate cholesteryl sulfate cholest-5-en-3beta-yl hydrogen sulfate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) o15118-1 NPC intracellular cholesterol transporter 1 Niemann-Pick C1 protein NPC1 protein protein human Homo sapiens Human MTARGLALGLLLLLLCPAQVFSQSCVWYGECGIAYGDKRYNCEYSGPPKPLPKDGYDLVQELCPGFFFGNVSLCCDVRQLQTLKDNLQLPLQFLSRCPSCFYNLLNLFCELTCSPRQSQFLNVTATEDYVDPVTNQTKTNVKELQYYVGQSFANAMYNACRDVEAPSSNDKALGLLCGKDADACNATNWIEYMFNKDNGQAPFTITPVFSDFPVHGMEPMNNATKGCDESVDEVTAPCSCQDCSIVCGPKPQPPPPPAPWTILGLDAMYVIMWITYMAFLLVFFGAFFAVWCYRKRYFVSEYTPIDSNIAFSVNASDKGEASCCDPVSAAFEGCLRRLFTRWGSFCVRNPGCVIFFSLVFITACSSGLVFVRVTTNPVDLWSAPSSQARLEKEYFDQHFGPFFRTEQLIIRAPLTDKHIYQPYPSGADVPFGPPLDIQILHQVLDLQIAIENITASYDNETVTLQDICLAPLSPYNTNCTILSVLNYFQNSHSVLDHKKGDDFFVYADYHTHFLYCVRAPASLNDTSLLHDPCLGTFGGPVFPWLVLGGYDDQNYNNATALVITFPVNNYYNDTEKLQRAQAWEKEFINFVKNYKNPNLTISFTAERSIEDELNRESDSDVFTVVISYAIMFLYISLALGHMKSCRRLLVDSKVSLGIAGILIVLSSVACSLGVFSYIGLPLTLIVIEVIPFLVLAVGVDNIFILVQAYQRDERLQGETLDQQLGRVLGEVAPSMFLSSFSETVAFFLGALSVMPAVHTFSLFAGLAVFIDFLLQITCFVSLLGLDIKRQEKNRLDIFCCVRGAEDGTSVQASESCLFRFFKNSYSPLLLKDWMRPIVIAIFVGVLSFSIAVLNKVDIGLDQSLSMPDDSYMVDYFKSISQYLHAGPPVYFVLEEGHDYTSSKGQNMVCGGMGCNNDSLVQQIFNAAQLDNYTRIGFAPSSWIDDYFDWVKPQSSCCRVDNITDQFCNASVVDPACVRCRPLTPEGKQRPQGGDFMRFLPMFLSDNPNPKCGKGGHAAYSSAVNILLGHGTRVGATYFMTYHTVLQTSADFIDALKKARLIASNVTETMGINGSAYRVFPYSVFYVFYEQYLTIIDDTIFNLGVSLGAIFLVTMVLLGCELWSAVIMCATIAMVLVNMFGVMWLWGISLNAVSLVNLVMSCGISVEFCSHITRAFTVSMKGSRVERAEEALAHMGSSVFSGITLTKFGGIVVLAFAKSQIFQIFYFRMYLAMVLLGATHGLIFLPVLLSYIGPSVNKAKSCATEERYKGTERERLLNF DA1523E09822E5C7 cholesterol sulfate-npc1-2 1 6 unspecified role unspecified role unspecified role unspecified role 7 unspecified role unspecified role neutral component neutral component 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 middle_lumenal_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role neutral component neutral component 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-293s Human 293S cell line Human 293s 293S cell line 12 15 direct interaction direct interaction Supp Table 1 2.4A R work =0.194 R free = 0.249 NAG is stated to be bound to NPC2 -Chain C/D (P61916) but as far as I can see this protein has been de-glycosylated -as stated in methods section, so NPC2 glycosylation would seem improbable.;so not entered. cholesterol sulfate-npc1-1 2 3 unspecified role unspecified role neutral component neutral component preprotrypsin signal sequence tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range sequence forming an antiparallel coiled coil tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range O35604:p.Phe503_Tyr504delinsAlaAla mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain FY AA Wed May 02 17:06:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe_tyr503-504ala_ala' Tue Jun 30 16:01:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe503_Tyr504delinsAlaAla' O35604:p.Asp502_Tyr504delinsAlaAlaAla mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain DFY AAA Wed May 02 17:29:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp_phe_tyr502-504ala_ala_ala' Tue Jun 30 19:28:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp502_Tyr504delinsAlaAlaAla' O35604:p.[Glu421Ala;Phe503_Tyr504delinsAlaAla] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain E A certain certain sequence position certain certain certain sequence position certain FY AA Mon Feb 27 13:46:52 GMT 2017 This feature has been corrected as a result of our quality control procedures. The original label was 'glu421ala, phe_tyr503-504ala_ala' Wed May 02 17:38:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu421ala,phe_tyr503-504ala_ala' Tue Jun 30 18:21:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Glu421Ala;Phe503_Tyr504delinsAlaAla]' n_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range middle_lumenal_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f 4 unspecified role unspecified role neutral component neutral component region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined bovin-milk Bos taurus milk milk 6 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Fig 3 2.4A Both the D502A,F503A,Y504A and E421F503A,Y504A mutant constructs gave a Kd of >3uM. I cannot enter the ">" into the parameter field.