bhf-ucl Neogenin interacts with hemojuvelin through its two membrane-proximal fibronectin type III domains. Neogenin interacts with hemojuvelin through its two membrane-proximal fibronectin type III domains. Biochemistry 2008 imex curation Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ., Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ. Cardiac - Interactions involving cardiac related proteins From methods: Human HJV and neogenin cDNAs; ectodomain of HJV (residues 1-401) with a C-terminal 6x-His tag. The longest neogenin ectodomain construct used DNA encoding the hydrophobic signal sequence (residues 1 – 33), the ectodomain (residues 34 to 1103), and a C-terminal 6x-His tag. Shorter constructs FNIII 5-6 and FNIII 1-6 were made the 1-33 signal sequence was kept, as well as the his tag and the ecodomain tail region was 1062-1103. In contrast, constructs 852-961 (FNIII 5) or 952-1103 (FNIII 6), have C-terminal 6x-His tag, and a gp67 hydrophobic signal. peptide. Plus: The shorter versions, sFNIII 5-6, sFNIII 5, and sFNIII 6, consisted of residues 853-1054, 853-952, and 952-1054, respectively. Recombinant proteins were purified from the supernatants of baculovirus-infected High Five (Hi-5) cells using Ni-NTA and gel filtration chromatography. Human BMP-2 was expressed in E. coli as inclusion bodies, purified and refolded Proteomic fragments of Neo1 regions based on UniProt record sequence with VDLFVI the N-terminal sequence of the peptide after Endoproteinase Glu-C digestion. This used in pull down expt. bjorkman@caltech.edu 15-Sep-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ., Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ. Cardiac - Interactions involving cardiac related proteins From methods: Human HJV and neogenin cDNAs; ectodomain of HJV (residues 1-401) with a C-terminal 6x-His tag. The longest neogenin ectodomain construct used DNA encoding the hydrophobic signal sequence (residues 1 – 33), the ectodomain (residues 34 to 1103), and a C-terminal 6x-His tag. Shorter constructs FNIII 5-6 and FNIII 1-6 were made the 1-33 signal sequence was kept, as well as the his tag and the ecodomain tail region was 1062-1103. In contrast, constructs 852-961 (FNIII 5) or 952-1103 (FNIII 6), have C-terminal 6x-His tag, and a gp67 hydrophobic signal. peptide. Plus: The shorter versions, sFNIII 5-6, sFNIII 5, and sFNIII 6, consisted of residues 853-1054, 853-952, and 952-1054, respectively. Recombinant proteins were purified from the supernatants of baculovirus-infected High Five (Hi-5) cells using Ni-NTA and gel filtration chromatography. Human BMP-2 was expressed in E. coli as inclusion bodies, purified and refolded Proteomic fragments of Neo1 regions based on UniProt record sequence with VDLFVI the N-terminal sequence of the peptide after Endoproteinase Glu-C digestion. This used in pull down expt. bjorkman@caltech.edu Accepted 2016-AUG-15 AT 10:59 BST AT 10:59 BST by ORCHARD Biochemistry 2008 Only protein-protein interactions imex curation imex curation Neogenin interacts with hemojuvelin through its two membrane-proximal fibronectin type III domains. Neogenin interacts with hemojuvelin through its two membrane-proximal fibronectin type III domains. Biochemistry 2008 imex curation Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ., Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ. Cardiac - Interactions involving cardiac related proteins From methods: Human HJV and neogenin cDNAs; ectodomain of HJV (residues 1-401) with a C-terminal 6x-His tag. The longest neogenin ectodomain construct used DNA encoding the hydrophobic signal sequence (residues 1 – 33), the ectodomain (residues 34 to 1103), and a C-terminal 6x-His tag. Shorter constructs FNIII 5-6 and FNIII 1-6 were made the 1-33 signal sequence was kept, as well as the his tag and the ecodomain tail region was 1062-1103. In contrast, constructs 852-961 (FNIII 5) or 952-1103 (FNIII 6), have C-terminal 6x-His tag, and a gp67 hydrophobic signal. peptide. Plus: The shorter versions, sFNIII 5-6, sFNIII 5, and sFNIII 6, consisted of residues 853-1054, 853-952, and 952-1054, respectively. Recombinant proteins were purified from the supernatants of baculovirus-infected High Five (Hi-5) cells using Ni-NTA and gel filtration chromatography. Human BMP-2 was expressed in E. coli as inclusion bodies, purified and refolded Proteomic fragments of Neo1 regions based on UniProt record sequence with VDLFVI the N-terminal sequence of the peptide after Endoproteinase Glu-C digestion. This used in pull down expt. bjorkman@caltech.edu 15-Sep-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro sls static light scattering sls predetermined predetermined participant predetermined Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ., Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ. Cardiac - Interactions involving cardiac related proteins From methods: Human HJV and neogenin cDNAs; ectodomain of HJV (residues 1-401) with a C-terminal 6x-His tag. The longest neogenin ectodomain construct used DNA encoding the hydrophobic signal sequence (residues 1 – 33), the ectodomain (residues 34 to 1103), and a C-terminal 6x-His tag. Shorter constructs FNIII 5-6 and FNIII 1-6 were made the 1-33 signal sequence was kept, as well as the his tag and the ecodomain tail region was 1062-1103. In contrast, constructs 852-961 (FNIII 5) or 952-1103 (FNIII 6), have C-terminal 6x-His tag, and a gp67 hydrophobic signal. peptide. Plus: The shorter versions, sFNIII 5-6, sFNIII 5, and sFNIII 6, consisted of residues 853-1054, 853-952, and 952-1054, respectively. Recombinant proteins were purified from the supernatants of baculovirus-infected High Five (Hi-5) cells using Ni-NTA and gel filtration chromatography. Human BMP-2 was expressed in E. coli as inclusion bodies, purified and refolded Proteomic fragments of Neo1 regions based on UniProt record sequence with VDLFVI the N-terminal sequence of the peptide after Endoproteinase Glu-C digestion. This used in pull down expt. bjorkman@caltech.edu Accepted 2016-AUG-15 AT 10:59 BST AT 10:59 BST by ORCHARD Biochemistry 2008 Only protein-protein interactions imex curation imex curation Neogenin interacts with hemojuvelin through its two membrane-proximal fibronectin type III domains. Neogenin interacts with hemojuvelin through its two membrane-proximal fibronectin type III domains. Biochemistry 2008 imex curation Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ., Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ. Cardiac - Interactions involving cardiac related proteins From methods: Human HJV and neogenin cDNAs; ectodomain of HJV (residues 1-401) with a C-terminal 6x-His tag. The longest neogenin ectodomain construct used DNA encoding the hydrophobic signal sequence (residues 1 – 33), the ectodomain (residues 34 to 1103), and a C-terminal 6x-His tag. Shorter constructs FNIII 5-6 and FNIII 1-6 were made the 1-33 signal sequence was kept, as well as the his tag and the ecodomain tail region was 1062-1103. In contrast, constructs 852-961 (FNIII 5) or 952-1103 (FNIII 6), have C-terminal 6x-His tag, and a gp67 hydrophobic signal. peptide. Plus: The shorter versions, sFNIII 5-6, sFNIII 5, and sFNIII 6, consisted of residues 853-1054, 853-952, and 952-1054, respectively. Recombinant proteins were purified from the supernatants of baculovirus-infected High Five (Hi-5) cells using Ni-NTA and gel filtration chromatography. Human BMP-2 was expressed in E. coli as inclusion bodies, purified and refolded Proteomic fragments of Neo1 regions based on UniProt record sequence with VDLFVI the N-terminal sequence of the peptide after Endoproteinase Glu-C digestion. This used in pull down expt. bjorkman@caltech.edu 15-Sep-2016: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ., Yang F., West AP., Allendorph GP., Choe S., Bjorkman PJ. Cardiac - Interactions involving cardiac related proteins From methods: Human HJV and neogenin cDNAs; ectodomain of HJV (residues 1-401) with a C-terminal 6x-His tag. The longest neogenin ectodomain construct used DNA encoding the hydrophobic signal sequence (residues 1 – 33), the ectodomain (residues 34 to 1103), and a C-terminal 6x-His tag. Shorter constructs FNIII 5-6 and FNIII 1-6 were made the 1-33 signal sequence was kept, as well as the his tag and the ecodomain tail region was 1062-1103. In contrast, constructs 852-961 (FNIII 5) or 952-1103 (FNIII 6), have C-terminal 6x-His tag, and a gp67 hydrophobic signal. peptide. Plus: The shorter versions, sFNIII 5-6, sFNIII 5, and sFNIII 6, consisted of residues 853-1054, 853-952, and 952-1054, respectively. Recombinant proteins were purified from the supernatants of baculovirus-infected High Five (Hi-5) cells using Ni-NTA and gel filtration chromatography. Human BMP-2 was expressed in E. coli as inclusion bodies, purified and refolded Proteomic fragments of Neo1 regions based on UniProt record sequence with VDLFVI the N-terminal sequence of the peptide after Endoproteinase Glu-C digestion. This used in pull down expt. bjorkman@caltech.edu Accepted 2016-AUG-15 AT 10:59 BST AT 10:59 BST by ORCHARD Biochemistry 2008 Only protein-protein interactions imex curation imex curation rgmc_human Hemojuvelin HJV HFE2 Hemochromatosis type 2 protein Hemojuvelin BMP coreceptor RGM domain family member C RGMC protein protein human Homo sapiens Human MGEPGQSPSPRSSHGSPPTLSTLTLLLLLCGHAHSQCKILRCNAEYVSSTLSLRGGGSSGALRGGGGGGRGGGVGSGGLCRALRSYALCTRRTARTCRGDLAFHSAVHGIEDLMIQHNCSRQGPTAPPPPRGPALPGAGSGLPAPDPCDYEGRFSRLHGRPPGFLHCASFGDPHVRSFHHHFHTCRVQGAWPLLDNDFLFVQATSSPMALGANATATRKLTIIFKNMQECIDQKVYQAEVDNLPVAFEDGSINGGDRPGGSSLSIQTANPGNHVEIQAAYIGTTIIIRQTAGQLSFSIKVAEDVAMAFSAEQDLQLCVGGCPPSQRLSRSERNRRGAITIDTARRLCKEGLPVEDAYFHSCVFDVLISGDPNFTVAAQAALEDARAFLPDLEKLHLFPSDAGVPLSSATLLAPLLSGLFVLWLCIQ 031C835F9B6DC06C bmp2_human Bone morphogenetic protein 2 BMP2 BMP2A Bone morphogenetic protein 2A protein protein human Homo sapiens Human MVAGTRCLLALLLPQVLLGGAAGLVPELGRRKFAAASSGRPSSQPSDEVLSEFELRLLSMFGLKQRPTPSRDAVVPPYMLDLYRRHSGQPGSPAPDHRLERAASRANTVRSFHHEESLEELPETSGKTTRRFFFNLSSIPTEEFITSAELQVFREQMQDALGNNSSFHHRINIYEIIKPATANSKFPVTRLLDTRLVNQNASRWESFDVTPAVMRWTAQGHANHGFVVEVAHLEEKQGVSKRHVRISRSLHQDEHSWSQIRPLLVTFGHDGKGHPLHKREKRQAKHKQRKRLKSSCKRHPLYVDFSDVGWNDWIVAPPGYHAFYCHGECPFPLADHLNSTNHAIVQTLVNSVNSKIPKACCVPTELSAISMLYLDENEKVVLKNYQDMVVEGCGCR DIP protein P12643 original sequence version: 173 DIP protein P12643 original sequence version: 172 DIP protein P12643 original sequence version: 159 DIP protein P12643 original sequence version: 178 20653A3987B25E60 neo1_human Neogenin NEO1 IGDCC2 NGN Immunoglobulin superfamily DCC subclass member 2 protein protein human Homo sapiens Human MAAERGARRLLSTPSFWLYCLLLLGRRAPGAAAARSGSAPQSPGASIRTFTPFYFLVEPVDTLSVRGSSVILNCSAYSEPSPKIEWKKDGTFLNLVSDDRRQLLPDGSLFISNVVHSKHNKPDEGYYQCVATVESLGTIISRTAKLIVAGLPRFTSQPEPSSVYAGNNAILNCEVNADLVPFVRWEQNRQPLLLDDRVIKLPSGMLVISNATEGDGGLYRCVVESGGPPKYSDEVELKVLPDPEVISDLVFLKQPSPLVRVIGQDVVLPCVASGLPTPTIKWMKNEEALDTESSERLVLLAGGSLEISDVTEDDAGTYFCIADNGNETIEAQAELTVQAQPEFLKQPTNIYAHESMDIVFECEVTGKPTPTVKWVKNGDMVIPSDYFKIVKEHNLQVLGLVKSDEGFYQCIAENDVGNAQAGAQLIILEHAPATTGPLPSAPRDVVASLVSTRFIKLTWRTPASDPHGDNLTYSVFYTKEGIARERVENTSHPGEMQVTIQNLMPATVYIFRVMAQNKHGSGESSAPLRVETQPEVQLPGPAPNLRAYAASPTSITVTWETPVSGNGEIQNYKLYYMEKGTDKEQDVDVSSHSYTINGLKKYTEYSFRVVAYNKHGPGVSTPDVAVRTLSDVPSAAPQNLSLEVRNSKSIMIHWQPPAPATQNGQITGYKIRYRKASRKSDVTETLVSGTQLSQLIEGLDRGTEYNFRVAALTINGTGPATDWLSAETFESDLDETRVPEVPSSLHVRPLVTSIVVSWTPPENQNIVVRGYAIGYGIGSPHAQTIKVDYKQRYYTIENLDPSSHYVITLKAFNNVGEGIPLYESAVTRPHTDTSEVDLFVINAPYTPVPDPTPMMPPVGVQASILSHDTIRITWADNSLPKHQKITDSRYYTVRWKTNIPANTKYKNANATTLSYLVTGLKPNTLYEFSVMVTKGRRSSTWSMTAHGTTFELVPTSPPKDVTVVSKEGKPKTIIVNWQPPSEANGKITGYIIYYSTDVNAEIHDWVIEPVVGNRLTHQIQELTLDTPYYFKIQARNSKGMGPMSEAVQFRTPKADSSDKMPNDQASGSGGKGSRLPDLGSDYKPPMSGSNSPHGSPTSPLDSNMLLVIIVSVGVITIVVVVIIAVFCTRRTTSHQKKKRAACKSVNGSHKYKGNSKDVKPPDLWIHHERLELKPIDKSPDPNPIMTDTPIPRNSQDITPVDNSMDSNIHQRRNSYRGHESEDSMSTLAGRRGMRPKMMMPFDSQPPQPVISAHPIHSLDNPHHHFHSSSLASPARSHLYHPGSPWPIGTSMSLSDRANSTESVRNTPSTDTMPASSSQTCCTDHQDPEGATSSSYLASSQEEDSGQSLPTAHVRPSHPLKSFAVPAIPPPGPPTYDPALPSTPLLSQQALNHHIHSVKTASIGTLGRSRPPMPVVVPSAPEVQETTRMLEDSESSYEPDELTKEMAHLEGLMKDLNAITTA DIP protein Q92859 original sequence version: 89 4AADF1EEBCAFD82C hjv-neo1-1 3 6 unspecified role unspecified role prey prey FNIII-6 fibronectin type III-like 6 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 4 unspecified role unspecified role bait bait purified purified c-terminal 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 Q6ZVN8:p.Asp172Ala asp172ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain D A Wed May 02 17:30:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp172ala' Tue Jun 30 18:56:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp172Ala' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction Table 1;Figure 2A, B hfe2-neo1-1 3 5 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role ancillary ancillary FNIII 5-6 fibronectin type III-like 5-6 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. I don't know how to add that in the results it states: After subtracting the residual response due to injecting a high concentration of FNIII 5-6 over the BMP-2 surface, we observed a higher maximal binding response for HJV in the presence of FNIII 5-6 than in its absence. Note that it also states no significant change in the derived affinity (KD = 67 nM) (Figure 4A). 4 unspecified role unspecified role prey prey c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. physical association physical association Figure 4 hfe2-neo1-2 1 6 unspecified role unspecified role prey prey FNIII 5-6 fibronectin type III-like 5-6 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 4 unspecified role unspecified role bait bait c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 25 29 direct interaction direct interaction Figure S1 bmp2-hfe2-1 3 5 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction Figure 4A hfe2-neo1-3 2 6 unspecified role unspecified role neutral component neutral component FNIII 5-6 fibronectin type III-like 5-6 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. 4 unspecified role unspecified role neutral component neutral component c-terminal c-terminal 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 trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction Figure 3