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/ http://www.ebi.ac.uk/ mihlan-2009-1 in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Mihlan M., Hebecker M., Dahse HM., Hälbich S., Huber-Lang M., Dahse R., Zipfel PF., Józsi M. CRP is composed of five identical subunits that are stabilized by calcium ions. This 125-kDa pentameric form (pCRP) is modified on calcium depletion, heat or urea treatment and dissociates into monomeric units of 23 kDa, termed monomeric CRP (mCRP). Expression system for CRP could not be ascertained from the publication and could be either serum or recombinant expression in E. coli mihaly.jozsi@hki-jena.de Molecular immunology 2009 Only protein-protein interactions imex curation imex curation mihlan-2009-2 in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Mihlan M., Hebecker M., Dahse HM., Hälbich S., Huber-Lang M., Dahse R., Zipfel PF., Józsi M. CRP is composed of five identical subunits that are stabilized by calcium ions. This 125-kDa pentameric form (pCRP) is modified on calcium depletion, heat or urea treatment and dissociates into monomeric units of 23 kDa, termed monomeric CRP (mCRP). Expression system for CRP could not be ascertained from the publication and could be either serum or recombinant expression in E. coli mihaly.jozsi@hki-jena.de Molecular immunology 2009 Only protein-protein interactions imex curation imex curation mihlan-2009-3 in vitro In vitro solid phase assay solid phase assay western blot western blot Immuno blot Immuno blot Mihlan M., Hebecker M., Dahse HM., Hälbich S., Huber-Lang M., Dahse R., Zipfel PF., Józsi M. CRP is composed of five identical subunits that are stabilized by calcium ions. This 125-kDa pentameric form (pCRP) is modified on calcium depletion, heat or urea treatment and dissociates into monomeric units of 23 kDa, termed monomeric CRP (mCRP). CFHR4A was incubated in test tubes with pCRP, then added to microtiter plate wells coated with PC-BSA. Bound proteins were eluted with SDS-sample buffer, separated by 10% SDS-PAGE and, after transfer onto a nitrocellulose membrane, they were detected with anti-CFHR4. Expression system for CRP could not be ascertained from the publication and could be either serum or recombinant expression in E. coli mihaly.jozsi@hki-jena.de Molecular immunology 2009 Only protein-protein interactions imex curation imex curation mihlan-2009-4 human-serum Homo sapiens serum Serum anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Mihlan M., Hebecker M., Dahse HM., Hälbich S., Huber-Lang M., Dahse R., Zipfel PF., Józsi M. CRP is composed of five identical subunits that are stabilized by calcium ions. This 125-kDa pentameric form (pCRP) is modified on calcium depletion, heat or urea treatment and dissociates into monomeric units of 23 kDa, termed monomeric CRP (mCRP). pCRP was precipitated from normal serum and sera of sepsis patients, by CRP-specific antibody. The precipitated proteins were separated by SDS-PAGE, transferred to membrane and analyzed using CFH antiserum. This anti-CFH antibody is stated to be goat anti-human CFH, purchased from Merck, and has an immunogen sequence HLVPDRKKDQYK. mihaly.jozsi@hki-jena.de Molecular immunology 2009 Only protein-protein interactions imex curation imex curation crp_human C-reactive protein CRP PTX1 protein protein human Homo sapiens Human MEKLLCFLVLTSLSHAFGQTDMSRKAFVFPKESDTSYVSLKAPLTKPLKAFTVCLHFYTELSSTRGYSIFSYATKRQDNEILIFWSKDIGYSFTVGGSEILFEVPEVTVAPVHICTSWESASGIVEFWVDGKPRVRKSLKKGYTVGAEASIILGQEQDSFGGNFEGSQSLVGDIGNVNMWDFVLSPDEINTIYLGGPFSPNVLNWRALKYEVQGEVFTKPQLWP q92496-1 Complement factor H-related protein 4 CFHR4 CFHL4 FHR4 FHR-4A protein protein human Homo sapiens Human MLLLINVILTLWVSCANGQEVKPCDFPEIQHGGLYYKSLRRLYFPAAAGQSYSYYCDQNFVTPSGSYWDYIHCTQDGWSPTVPCLRTCSKSDVEIENGFISESSSIYILNEETQYNCKPGYATAEGNSSGSITCLQNGWSTQPICIKFCDMPVFENSRAKSNGMWFKLHDTLDYECYDGYESSYGNTTDSIVCGEDGWSHLPTCYNSSENCGPPPPISNGDTTSFPQKVYLPWSRVEYQCQSYYELQGSKYVTCSNGDWSEPPRCISMKPCEFPEIQHGHLYYENTRRPYFPVATGQSYSYYCDQNFVTPSGSYWDYIHCTQDGWLPTVPCLRTCSKSDIEIENGFISESSSIYILNKEIQYKCKPGYATADGNSSGSITCLQNGWSAQPICIKFCDMPVFENSRAKSNGMRFKLHDTLDYECYDGYEISYGNTTGSIVCGEDGWSHFPTCYNSSEKCGPPPPISNGDTTSFLLKVYVPQSRVEYQCQSYYELQGSNYVTCSNGEWSEPPRCIHPCIITEENMNKNNIQLKGKSDIKYYAKTGDTIEFMCKLGYNANTSVLSFQAVCREGIVEYPRCE q92496-3 Complement factor H-related protein 4 CFHR4 CFHL4 FHR4 FHR-4B protein protein human Homo sapiens Human MLLLINVILTLWVSCANGQEVKPCDFPEIQHGGLYYKSLRRLYFPAAAGQSYSYYCDQNFVTPSGSYWDYIHCTQDGWSPTVPCLRTCSKSDIEIENGFISESSSIYILNKEIQYKCKPGYATADGNSSGSITCLQNGWSAQPICIKFCDMPVFENSRAKSNGMRFKLHDTLDYECYDGYEISYGNTTGSIVCGEDGWSHFPTCYNSSEKCGPPPPISNGDTTSFLLKVYVPQSRVEYQCQSYYELQGSNYVTCSNGEWSEPPRCIHPCIITEENMNKNNIQLKGKSDIKYYAKTGDTIEFMCKLGYNANTSVLSFQAVCREGIVEYPRCE cfah_human Complement factor H CFH HF HF1 HF2 H factor 1 protein protein human Homo sapiens Human MRLLAKIICLMLWAICVAEDCNELPPRRNTEILTGSWSDQTYPEGTQAIYKCRPGYRSLGNVIMVCRKGEWVALNPLRKCQKRPCGHPGDTPFGTFTLTGGNVFEYGVKAVYTCNEGYQLLGEINYRECDTDGWTNDIPICEVVKCLPVTAPENGKIVSSAMEPDREYHFGQAVRFVCNSGYKIEGDEEMHCSDDGFWSKEKPKCVEISCKSPDVINGSPISQKIIYKENERFQYKCNMGYEYSERGDAVCTESGWRPLPSCEEKSCDNPYIPNGDYSPLRIKHRTGDEITYQCRNGFYPATRGNTAKCTSTGWIPAPRCTLKPCDYPDIKHGGLYHENMRRPYFPVAVGKYYSYYCDEHFETPSGSYWDHIHCTQDGWSPAVPCLRKCYFPYLENGYNQNYGRKFVQGKSIDVACHPGYALPKAQTTVTCMENGWSPTPRCIRVKTCSKSSIDIENGFISESQYTYALKEKAKYQCKLGYVTADGETSGSITCGKDGWSAQPTCIKSCDIPVFMNARTKNDFTWFKLNDTLDYECHDGYESNTGSTTGSIVCGYNGWSDLPICYERECELPKIDVHLVPDRKKDQYKVGEVLKFSCKPGFTIVGPNSVQCYHFGLSPDLPICKEQVQSCGPPPELLNGNVKEKTKEEYGHSEVVEYYCNPRFLMKGPNKIQCVDGEWTTLPVCIVEESTCGDIPELEHGWAQLSSPPYYYGDSVEFNCSESFTMIGHRSITCIHGVWTQLPQCVAIDKLKKCKSSNLIILEEHLKNKKEFDHNSNIRYRCRGKEGWIHTVCINGRWDPEVNCSMAQIQLCPPPPQIPNSHNMTTTLNYRDGEKVSVLCQENYLIQEGEEITCKDGRWQSIPLCVEKIPCSQPPQIEHGTINSSRSSQESYAHGTKLSYTCEGGFRISEENETTCYMGKWSSPPQCEGLPCKSPPEISHGVVAHMSDSYQYGEEVTYKCFEGFGIDGPAIAKCLGEKWSHPPSCIKTDCLSLPSFENAIPMGEKKDVYKAGEQVTYTCATYYKMDGASNVTCINSRWTGRPTCRDTSCVNPPTVQNAYIVSRQMSKYPSGERVRYQCRSPYEMFGDEEVMCLNGNWTEPPQCKDSTGKCGPPPPIDNGDITSFPLSVYAPASSVEYQCQNLYQLEGNKRITCRNGQWSEPPKCLHPCVISREIMENYNIALRWTAKQKLYSRTGESVEFVCKRGYRLSSRSHTLRTTCWDGKLEYPTCAKR albu_bovin Albumin ALB BSA protein protein bovin Bos taurus (Bovine) Bovine MKWVTFISLLLLFSSAYSRGVFRRDTHKSEIAHRFKDLGEEHFKGLVLIAFSQYLQQCPFDEHVKLVNELTEFAKTCVADESHAGCEKSLHTLFGDELCKVASLRETYGDMADCCEKQEPERNECFLSHKDDSPDLPKLKPDPNTLCDEFKADEKKFWGKYLYEIARRHPYFYAPELLYYANKYNGVFQECCQAEDKGACLLPKIETMREKVLASSARQRLRCASIQKFGERALKAWSVARLSQKFPKAEFVEVTKLVTDLTKVHKECCHGDLLECADDRADLAKYICDNQDTISSKLKECCDKPLLEKSHCIAEVEKDAIPENLPPLTADFAEDKDVCKNYQEAKDAFLGSFLYEYSRRHPEYAVSVLLRLAKEYEATLEECCAKDDPHACYSTVFDKLKHLVDEPQNLIKQNCDQFEKLGEYGFQNALIVRYTRKVPQVSTPTLVEVSRSLGKVGTRCCTKPESERMPCTEDYLSLILNRLCVLHEKTPVSEKVTKCCTESLVNRRPCFSALTPDETYVPKAFDEKLFTFHADICTLPDTEKQIKKQTALVELLKHKPKATEEQLKTVMENFVAFVDKCCAADDKEACFAVEGPKLVVSTQTALA phosphocholine phosphocholine Choline phosphate ChoP N-Trimethyl-2-aminoethylphosphonate O-Phosphocholine O-phosphocholine PHOSPHOCHOLINE Phosphocholine Phosphoryl-choline Phosphorylcholine Trimethyl(2-(phosphonooxy)ethyl)ammonium N,N,N-trimethyl-2-(phosphonooxy)ethanaminium small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cfhr4-crp-5 3595086 N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 5.0 N/A 3595091 unspecified role unspecified role bait bait 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 false SCR/Sushi_domains 5-9 binding region binding-associated region binding region range range certain certain sequence position certain false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction false false false 3 A/B-1,4 B/D Ca2+ The somewhat stronger binding of mCRP to CFHR4A "5-9" than to CFHR4A and CFHR4B is unexplained. cfhr4-crp-1 3595086 N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 5.0 N/A 3595092 unspecified role unspecified role bait bait purified purified 9inse 9inse direct interaction direct interaction false false false 3 A/B-2,4 B/D Ca2+ cfhr4-crp-2 3595086 N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 N/A 3595091 unspecified role unspecified role bait bait 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 false SCR/Sushi_domains 5-9 binding region binding-associated region binding region range range certain certain sequence position certain false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction false false false 3 A/B-3 Ca2+ mCRP bound only very weakly to the CFHR4 isoforms. The somewhat stronger binding of mCRP to CFHR4A "5-9" than to CFHR4A and CFHR4B is unexplained. cfhr4-crp-3 3595086 N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 N/A 3595092 unspecified role unspecified role bait bait purified purified 9inse 9inse direct interaction direct interaction false false false 3 A/B-4 Ca2+ mCRP bound only very weakly to the CFHR4 isoforms. cfhr4-crp-4 3595086 N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 N/A 3595093 unspecified role unspecified role bait bait purified purified human-serum Homo sapiens serum Serum direct interaction direct interaction false false false 3 A/B-5 mCRP bound to CFH in a calcium-independent manner. cfhr4-crp-6 3595086 N/A 3595090 unspecified role unspecified role bait bait purified purified Stoichiometry: 5.0 N/A 3595091 unspecified role unspecified role prey prey 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 false SCR/Sushi_domains 5-9 binding region binding-associated region binding region range range certain certain sequence position certain false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction false false false 3 C/D-1, 5A Ca2+ cfhr4-crp-7 3595086 N/A 3595090 unspecified role unspecified role bait bait purified purified Stoichiometry: 5.0 N/A 3595092 unspecified role unspecified role prey prey purified purified 9inse 9inse direct interaction direct interaction false false false 3 C/D-2 Ca2+ cfh-crp-4 3595086 N/A 3595090 unspecified role unspecified role bait bait purified purified Stoichiometry: 1.0 N/A 3595093 unspecified role unspecified role bait bait purified purified human-serum Homo sapiens serum Serum direct interaction direct interaction false false false 3 C/D-3 mCRP bound to CFH in a calcium-independent manner. cfh-7 3595086 N/A 3595090 unspecified role unspecified role bait bait purified purified Stoichiometry: 5.0 N/A 3595093 unspecified role unspecified role bait bait purified purified human-serum Homo sapiens serum Serum direct interaction direct interaction false false false 3 C/D-4 pCRP bound to CFH in a calcium-independent manner. cfhr4-crp-10 3595086 N/A 3595090 competitor competitor competitor unspecified role unspecified role purified purified N/A 3595090 unspecified role unspecified role bait bait purified purified Stoichiometry: 5.0 N/A 3595091 unspecified role unspecified role prey prey 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 false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction false false false 5B Ca2+ The somewhat stronger binding of mCRP to CFHR4A "5-9" than to CFHR4A and CFHR4B is unexplained. cfhr4-crp-8 3595087 N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 5.0 N/A 3595092 unspecified role unspecified role bait bait purified purified 9inse 9inse direct interaction direct interaction false false false 3 E/F Ca2+; in the absence of calcium a fast dissociation was observed cfhr4-crp-9 3595087 N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 5.0 N/A 3595091 unspecified role unspecified role bait bait 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 false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction false false false 3 E/F, 4C Ca2+; in the absence of calcium a fast dissociation was observed cfhr4-crp-11 3595088 N/A 3595094 unspecified role unspecified role bait bait purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false bovin-serum Bovine serum serum Serum N/A 3595090 unspecified role unspecified role prey prey purified purified Stoichiometry: 5.0 N/A 3595091 unspecified role unspecified role prey prey 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 false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection N/A 3595095 unspecified role unspecified role bait bait whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) 3595148 3595143 physical association physical association false false false 5C crp-cfh-3 3595089 N/A 3595093 unspecified role unspecified role prey prey N/A 3595090 unspecified role unspecified role bait bait physical association physical association false false false 5D It is stated that CFHR4 was detected in the sera of sepsis patients using anti-CFH antisera, but it is unclear how this can be possible, as the immunogen sequence for the antibody used (HLVPDRKKDQYK) is not found even partially in the CFHR4 sequence. However there is an extra band present in the sera from sepsis patients which is not present in normal sera.