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/ clark-2014-1 in vitro In vitro solid phase assay solid phase assay predetermined predetermined participant predetermined Clark SJ., Schmidt CQ., White AM., Hakobyan S., Morgan BP., Bishop PN. Heparin, Heparan sulfate or selectively desulfated derivatives of Heparin: 2-O desulfated (2-O deS); 6-O desulfated (6-O deS) and N-desulfated (N-deS) were used. simon.clark-3@manchester.ac.uk Journal of immunology (Baltimore, Md. : 1950) 2014 Only protein-protein interactions imex curation imex curation clark-2014-2 in vitro In vitro pull down pull down predetermined predetermined participant predetermined Clark SJ., Schmidt CQ., White AM., Hakobyan S., Morgan BP., Bishop PN. simon.clark-3@manchester.ac.uk Journal of immunology (Baltimore, Md. : 1950) 2014 Only protein-protein interactions imex curation imex curation clark-2014-3 in vitro In vitro protease assay protease assay predetermined predetermined participant predetermined Clark SJ., Schmidt CQ., White AM., Hakobyan S., Morgan BP., Bishop PN. simon.clark-3@manchester.ac.uk Journal of immunology (Baltimore, Md. : 1950) 2014 Only protein-protein interactions imex curation imex curation p08603-2 Complement factor H CFH HF HF1 HF2 FHL-1 H factor 1 protein protein human Homo sapiens Human MRLLAKIICLMLWAICVAEDCNELPPRRNTEILTGSWSDQTYPEGTQAIYKCRPGYRSLGNVIMVCRKGEWVALNPLRKCQKRPCGHPGDTPFGTFTLTGGNVFEYGVKAVYTCNEGYQLLGEINYRECDTDGWTNDIPICEVVKCLPVTAPENGKIVSSAMEPDREYHFGQAVRFVCNSGYKIEGDEEMHCSDDGFWSKEKPKCVEISCKSPDVINGSPISQKIIYKENERFQYKCNMGYEYSERGDAVCTESGWRPLPSCEEKSCDNPYIPNGDYSPLRIKHRTGDEITYQCRNGFYPATRGNTAKCTSTGWIPAPRCTLKPCDYPDIKHGGLYHENMRRPYFPVAVGKYYSYYCDEHFETPSGSYWDHIHCTQDGWSPAVPCLRKCYFPYLENGYNQNYGRKFVQGKSIDVACHPGYALPKAQTTVTCMENGWSPTPRCIRVSFTL heparin Heparin small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) heparan_sulfate Heparan_Sulfate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) cfai_human Complement factor I CFI IF C3B/C4B inactivator protein protein human Homo sapiens Human MKLLHVFLLFLCFHLRFCKVTYTSQEDLVEKKCLAKKYTHLSCDKVFCQPWQRCIEGTCVCKLPYQCPKNGTAVCATNRRSFPTYCQQKSLECLHPGTKFLNNGTCTAEGKFSVSLKHGNTDSEGIVEVKLVDQDKTMFICKSSWSMREANVACLDLGFQQGADTQRRFKLSDLSINSTECLHVHCRGLETSLAECTFTKRRTMGYQDFADVVCYTQKADSPMDDFFQCVNGKYISQMKACDGINDCGDQSDELCCKACQGKGFHCKSGVCIPSQYQCNGEVDCITGEDEVGCAGFASVTQEETEILTADMDAERRRIKSLLPKLSCGVKNRMHIRRKRIVGGKRAQLGDLPWQVAIKDASGITCGGIYIGGCWILTAAHCLRASKTHRYQIWTTVVDWIHPDLKRIVIEYVDRIIFHENYNAGTYQNDIALIEMKKDGNKKDCELPRSIPACVPWSPYLFQPNDTCIVSGWGREKDNERVFSLQWGEVKLISNCSKFYGNRFYEKEMECAGTYDGSIDACKGDSGGPLVCMDANNVTYVWGVVSWGENCGKPEFPGVYTKVANYFDWISYHVGRPFISQYNV p01024-pro_0000005911 Complement C3b alpha' chain C3 CPAMD1 C3 and PZP-like alpha-2-macroglobulin domain-containing protein 1 protein protein human Homo sapiens Human SNLDEDIIAEENIVSRSEFPESWLWNVEDLKEPPKNGISTKLMNIFLKDSITTWEILAVSMSDKKGICVADPFEVTVMQDFFIDLRLPYSVVRNEQVEIRAVLYNYRQNQELKVRVELLHNPAFCSLATTKRRHQQTVTIPPKSSLSVPYVIVPLKTGLQEVEVKAAVYHHFISDGVRKSLKVVPEGIRMNKTVAVRTLDPERLGREGVQKEDIPPADLSDQVPDTESETRILLQGTPVAQMTEDAVDAERLKHLIVTPSGCGEQNMIGMTPTVIAVHYLDETEQWEKFGLEKRQGALELIKKGYTQQLAFRQPSSAFAAFVKRAPSTWLTAYVVKVFSLAVNLIAIDSQVLCGAVKWLILEKQKPDGVFQEDAPVIHQEMIGGLRNNNEKDMALTAFVLISLQEAKDICEEQVNSLPGSITKAGDFLEANYMNLQRSYTVAIAGYALAQMGRLKGPLLNKFLTTAKDKNRWEDPGKQLYNVEATSYALLALLQLKDFDFVPPVVRWLNEQRYYGGGYGSTQATFMVFQALAQYQKDAPDHQELNLDVSLQLPSRSSKITHRIHWESASLLRSEETKENEGFTVTAEGKGQGTLSVVTMYHAKAKDQLTCNKFDLKVTIKPAPETEKRPQDAKNTMILEICTRYRGDQDATMSILDISMMTGFAPDTDDLKQLANGVDRYISKYELDKAFSDRNTLIIYLDKVSHSEDDCLAFKVHQYFNVELIQPGAVKVYAYYNLEESCTRFYHPEKEDGKLNKLCRDELCRCAEENCFIQKSDDKVTLEERLDKACEPGVDYVYKTRLVKVQLSNDFDEYIMAIEQTIKSGSDEVQVGQQRTFISPIKCREALKLEEKKHYLMWGLSSDFWGEKPNLSYIIGKDTWVEHWPEEDECQDEENQKQCQDLGAFTESMVVFGCPN 749 1663 heparin-cfh-5 3631544 N/A 3631547 unspecified role unspecified role prey prey purified purified region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined certain certain sequence position certain false 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 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 false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3631548 unspecified role unspecified role bait bait purified purified desulphated_residues carbohydrate chemical modification with no effect carbohydrate chemical modification with no effect carbohydrate chemical modification not affecting interaction undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false pig Sus scrofa (Pig) Pig direct interaction direct interaction false false false 4B Fully sulphated Heparin bound both the Y402 and H402 variants. However desulphation at the 6-O, 2-O and N-positions , significantly reduced the binding of the AMD-associated 402H form to FHL-1. heparin-cfh-6 3631544 N/A 3631547 unspecified role unspecified role prey prey purified purified P08603-2:p.Tyr402His 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 false region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined certain certain sequence position certain false 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 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 false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3631548 unspecified role unspecified role bait bait purified purified desulphated_residues carbohydrate chemical modification decreasing strength carbohydrate chemical modification decreasing interaction strength carbohydrate chemical modification decreasing strength undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false pig Sus scrofa (Pig) Pig direct interaction direct interaction false false false 4C Fully sulphated Heparin bound both the Y402 and H402 variants. However desulphation at the 6-O, 2-O and N-positions , significantly reduced the binding of the AMD-associated 402H form to FHL-1. heparin-cfh-7 3631544 N/A 3631547 unspecified role unspecified role prey prey purified purified P08603-2:p.Tyr402His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined certain certain sequence position certain false 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 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 false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3631549 unspecified role unspecified role bait bait purified purified pig Sus scrofa (Pig) Pig direct interaction direct interaction false false false 4E Heparan sulfate contains large regions with low sulfated disaccharides and overall has much less sulfation than heparin. heparan_sulfate-cfh-1 3631544 N/A 3631547 unspecified role unspecified role prey prey purified purified P08603-2:p.Tyr402His mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined certain certain sequence position certain false 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 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 false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3631549 unspecified role unspecified role bait bait purified purified bovin-kidney bovin kidney kidney kidney [Renal] direct interaction direct interaction false false false 4F Heparan sulfate contains large regions with low sulfated disaccharides and overall has much less sulfation than heparin heparin-cfh-4 3631544 N/A 3631547 unspecified role unspecified role prey prey purified purified region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined certain certain sequence position certain false 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 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 false P08603-2:p.Tyr402His 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 false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3631548 unspecified role unspecified role bait bait purified purified pig Sus scrofa (Pig) Pig direct interaction direct interaction false false false 4B/C Fully sulphated Heparin bound both the Y402 and H402 variants. However desulphation at the 6-O, 2-O and N-positions , significantly reduced the binding of the AMD-associated 402H form to FHL-1. heparin-cfh-8 3631545 N/A 3631547 unspecified role unspecified role prey prey purified purified P08603-2:p.Tyr402His mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined certain certain sequence position certain false 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 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 false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3631548 unspecified role unspecified role bait bait purified purified pig Sus scrofa (Pig) Pig direct interaction direct interaction false false false Supp. Fig 2 The 402H form of FHL-1 bound better than the 402Y form to highly sulfated heparin. c3-cfh-22 3631546 N/A 3631547 cofactor cofactor coenzyme neutral component neutral component purified purified P08603-2:p.Tyr402His 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 false region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined certain certain sequence position certain false 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 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 false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3631550 enzyme enzyme neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3631551 enzyme target enzyme target substrate substrate neutral component neutral component purified purified human-blood human blood blood BLOOD protein cleavage protein cleavage false false false Supp. Fig 3