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/ Complement factor H binds at two independent sites to C-reactive protein in acute phase concentrations. Complement factor H binds at two independent sites to C-reactive protein in acute phase concentrations. The Journal of biological chemistry 2010 imex curation Okemefuna AI., Nan R., Miller A., Gor J., Perkins SJ. 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. The origin of Human CRP was from "malignant effusion fluids" notably pleural effusion or ascites. s.perkins@medsch.ucl.ac.uk 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro saxs x ray scattering saxs predetermined predetermined participant predetermined Okemefuna AI., Nan R., Miller A., Gor J., Perkins SJ. 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. s.perkins@medsch.ucl.ac.uk Accepted 2019-NOV-13 AT 17:00 GMT AT 17:00 GMT by PPORRAS The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation Complement factor H binds at two independent sites to C-reactive protein in acute phase concentrations. Complement factor H binds at two independent sites to C-reactive protein in acute phase concentrations. The Journal of biological chemistry 2010 imex curation Okemefuna AI., Nan R., Miller A., Gor J., Perkins SJ. 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. The origin of Human CRP was from "malignant effusion fluids" notably pleural effusion or ascites. s.perkins@medsch.ucl.ac.uk 21-Nov-2019: 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 Okemefuna AI., Nan R., Miller A., Gor J., Perkins SJ. 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. I have not entered stoichiometry as stated in text for most SPRs as they state 1:1 and various other figures for Fig 3/4, but I do not know when the "1" refers to the CRP pentamer, and when not, so no values entered, apart from CRP =5. s.perkins@medsch.ucl.ac.uk Accepted 2019-NOV-13 AT 17:00 GMT AT 17:00 GMT by PPORRAS The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation Complement factor H binds at two independent sites to C-reactive protein in acute phase concentrations. Complement factor H binds at two independent sites to C-reactive protein in acute phase concentrations. The Journal of biological chemistry 2010 imex curation Okemefuna AI., Nan R., Miller A., Gor J., Perkins SJ. 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. The origin of Human CRP was from "malignant effusion fluids" notably pleural effusion or ascites. s.perkins@medsch.ucl.ac.uk 21-Nov-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Okemefuna AI., Nan R., Miller A., Gor J., Perkins SJ. 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. s.perkins@medsch.ucl.ac.uk Accepted 2019-NOV-13 AT 17:00 GMT AT 17:00 GMT by PPORRAS The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation crp_human C-reactive protein CRP PTX1 protein protein human Homo sapiens Human MEKLLCFLVLTSLSHAFGQTDMSRKAFVFPKESDTSYVSLKAPLTKPLKAFTVCLHFYTELSSTRGYSIFSYATKRQDNEILIFWSKDIGYSFTVGGSEILFEVPEVTVAPVHICTSWESASGIVEFWVDGKPRVRKSLKKGYTVGAEASIILGQEQDSFGGNFEGSQSLVGDIGNVNMWDFVLSPDEINTIYLGGPFSPNVLNWRALKYEVQGEVFTKPQLWP 669228636A8544F6 cfah_human Complement factor H CFH HF HF1 HF2 H factor 1 protein protein human Homo sapiens Human MRLLAKIICLMLWAICVAEDCNELPPRRNTEILTGSWSDQTYPEGTQAIYKCRPGYRSLGNVIMVCRKGEWVALNPLRKCQKRPCGHPGDTPFGTFTLTGGNVFEYGVKAVYTCNEGYQLLGEINYRECDTDGWTNDIPICEVVKCLPVTAPENGKIVSSAMEPDREYHFGQAVRFVCNSGYKIEGDEEMHCSDDGFWSKEKPKCVEISCKSPDVINGSPISQKIIYKENERFQYKCNMGYEYSERGDAVCTESGWRPLPSCEEKSCDNPYIPNGDYSPLRIKHRTGDEITYQCRNGFYPATRGNTAKCTSTGWIPAPRCTLKPCDYPDIKHGGLYHENMRRPYFPVAVGKYYSYYCDEHFETPSGSYWDHIHCTQDGWSPAVPCLRKCYFPYLENGYNQNYGRKFVQGKSIDVACHPGYALPKAQTTVTCMENGWSPTPRCIRVKTCSKSSIDIENGFISESQYTYALKEKAKYQCKLGYVTADGETSGSITCGKDGWSAQPTCIKSCDIPVFMNARTKNDFTWFKLNDTLDYECHDGYESNTGSTTGSIVCGYNGWSDLPICYERECELPKIDVHLVPDRKKDQYKVGEVLKFSCKPGFTIVGPNSVQCYHFGLSPDLPICKEQVQSCGPPPELLNGNVKEKTKEEYGHSEVVEYYCNPRFLMKGPNKIQCVDGEWTTLPVCIVEESTCGDIPELEHGWAQLSSPPYYYGDSVEFNCSESFTMIGHRSITCIHGVWTQLPQCVAIDKLKKCKSSNLIILEEHLKNKKEFDHNSNIRYRCRGKEGWIHTVCINGRWDPEVNCSMAQIQLCPPPPQIPNSHNMTTTLNYRDGEKVSVLCQENYLIQEGEEITCKDGRWQSIPLCVEKIPCSQPPQIEHGTINSSRSSQESYAHGTKLSYTCEGGFRISEENETTCYMGKWSSPPQCEGLPCKSPPEISHGVVAHMSDSYQYGEEVTYKCFEGFGIDGPAIAKCLGEKWSHPPSCIKTDCLSLPSFENAIPMGEKKDVYKAGEQVTYTCATYYKMDGASNVTCINSRWTGRPTCRDTSCVNPPTVQNAYIVSRQMSKYPSGERVRYQCRSPYEMFGDEEVMCLNGNWTEPPQCKDSTGKCGPPPPIDNGDITSFPLSVYAPASSVEYQCQNLYQLEGNKRITCRNGQWSEPPKCLHPCVISREIMENYNIALRWTAKQKLYSRTGESVEFVCKRGYRLSSRSHTLRTTCWDGKLEYPTCAKR DIP protein P08603 original sequence version: 161 DIP protein P08603-1 original sequence version: 149 3C26D62A2BF9BFEE cfh-23 3 5 unspecified role unspecified role neutral component neutral component purified purified human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction 2A/B, peak 2 NaCl 137mM cfh-22 3 5 unspecified role unspecified role neutral component neutral component purified purified human-plasma homo sapiens blood plasma plasma blood plasma 5 unspecified role unspecified role neutral component neutral component purified purified human-plasma homo sapiens blood plasma plasma blood plasma direct interaction direct interaction 2A/B, peaks 3-8 NaCl 137mM cfh-24 3 4 unspecified role unspecified role neutral component neutral component purified purified human Homo sapiens Human direct interaction direct interaction 2A/B, peak P it does not look to me that 137mM NaCl is inhibiting here crp-35 3 4 unspecified role unspecified role neutral component neutral component purified purified human Homo sapiens Human direct interaction direct interaction 2A/B, peak D NaCl 137mM crp-36 3 5 unspecified role unspecified role neutral component neutral component purified purified human-plasma homo sapiens blood plasma plasma blood plasma 4 unspecified role unspecified role neutral component neutral component purified purified human Homo sapiens Human direct interaction direct interaction 2A/B, peak A 137mM NaCl At higher CRP concentrations, peak C is attributed to the FH·CRP2 complex. Peak B may correspond to a stoichiometric 1:1 complex between FH and CRP. the text makes it obvious they do not know stoichiometry for Peaks B/C, nor what the peaks contain, so have just entered a comment. cfh-crp-22 2 5 unspecified role unspecified role bait bait purified purified human-plasma homo sapiens blood plasma plasma blood plasma 4 unspecified role unspecified role prey prey purified purified human Homo sapiens Human direct interaction direct interaction 3 A-D 137mM NaCl In 50 mm NaCl CRP aggregated onto immobilized FH;using 137 mm NaCl, CRP binding was greatly weakened. cfh-crp-23 2 5 unspecified role unspecified role prey prey purified purified human-plasma homo sapiens blood plasma plasma blood plasma 4 unspecified role unspecified role bait bait purified purified human Homo sapiens Human direct interaction direct interaction 3 E, F 137mM NaCl In 50 mm NaCl CRP aggregated onto immobilized FH;using 137 mm NaCl, CRP binding was greatly weakened. Figure 3E -50 mM NaCl: KD values are estimates only. Two apparent equilibrium constants, KD1-app and KD2-app, of 3.4 and 0.4 um, respectively, were obtained. Figure 3F -137nM NaCl:the FH-CRP interaction was weaker and proceeded to steady state equilibrium, from which an equilibrium KD value of 4.2 um was determined. Fig 3F states "This analysis was well represented by a 1:1 interaction." however they do not confirm if this is the CRP pentamer + 1 mol FH, so have not entered this. crp-cfh-35 2 5 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 SCR/Sushi_domains 16-20 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain C-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range picpa Pichia pastoris (Yeast) Yeast Pichia pastoris 4 unspecified role unspecified role bait bait purified purified human Homo sapiens Human direct interaction direct interaction 4 A/B/E 137mM NaCl The KD value for SCR-16/20 was determined to be 15.3 um, at NaCl 137mM. crp-cfh-36 2 5 unspecified role unspecified role prey prey purified purified SCR/Sushi_domains 6-8 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 P08603:p.Tyr402His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Y H Tue Jun 30 16:53:19 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr402His' ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role bait bait purified purified human Homo sapiens Human direct interaction direct interaction 4 C/D/F Details of SCR 6-8 construct from Ref. 34. The FH Tyr-402 allotype was bound in similar amounts in 50 and 137 mm NaCl, whereas His-402 allotype binding was mostly absent in 50 mm NaCl. cfh-crp-24 1 5 unspecified role unspecified role neutral component neutral component purified purified human-plasma homo sapiens blood plasma plasma blood plasma 4 unspecified role unspecified role neutral component neutral component purified purified human Homo sapiens Human direct interaction direct interaction 5 137mM NaCl