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/ salinas-2014-7 in vitro In vitro solid phase assay solid phase assay tag fluorescence tag visualisation by fluorescence tag fluorescence Salinas ND., Paing MM., Tolia NH. terminal sialic acid was removed by neuraminidase treatment. GPA (ecd):- Sequence, as shown in Figure 1A appears to be that of GenBank: AAA52624.1. This sequence has been remapped to P02724 due to 'Erroneous initiation' of AAA52624.1. EBA-175 P. falciparum protein:- Ref 7 states this is a protein of 616 amino acid and PDBe entries give UniProt ID =Q25735 for the protein. No strain of the parasite is stated, and Q25735 has been remapped to reviewed entry P19214 (aas 145-760) with which it has 99% homology. tolia@wustl.edu mBio 2014 imex curation Only protein-protein interactions imex curation salinas-2014-8 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Salinas ND., Paing MM., Tolia NH. GPA (ecd):- Sequence, as shown in Figure 1A appears to be that of GenBank: AAA52624.1. This sequence has been remapped to P02724 due to 'Erroneous initiation' of AAA52624.1. EBA-175 P. falciparum protein:- Ref 7 states this is a protein of 616 amino acid and PDBe entries give UniProt ID =Q25735 for the protein. No strain of the parasite is stated, and Q25735 has been remapped to reviewed entry P19214 (aas 145-760) with which it has 99% homology. tolia@wustl.edu mBio 2014 imex curation Only protein-protein interactions imex curation lec_soybn Lectin LE1 Agglutinin SBA protein protein soybn Glycine max (Soybean) Glycine hispida MATSKLKTQNVVVSLSLTLTLVLVLLTSKANSAETVSFSWNKFVPKQPNMILQGDAIVTSSGKLQLNKVDENGTPKPSSLGRALYSTPIHIWDKETGSVASFAASFNFTFYAPDTKRLADGLAFFLAPIDTKPQTHAGYLGLFNENESGDQVVAVEFDTFRNSWDPPNPHIGINVNSIRSIKTTSWDLANNKVAKVLITYDASTSLLVASLVYPSQRTSNILSDVVDLKTSLPEWVRIGFSAATGLDIPGESHDVLSWSFASNLPHASSNIDPLDLTSFVLHEAI glpa_human Glycophorin-A PAS-2 Sialoglycoprotein alpha MN sialoglycoprotein GYPA GPA protein protein human Homo sapiens Human MYGKIIFVLLLSEIVSISASSTTGVAMHTSTSSSVTKSYISSQTNDTHKRDTYAATPRAHEVSEISVRTVYPPEEETGERVQLAHHFSEPEITLIIFGVMAGVIGTILLISYGIRRLIKKSPSDVKPLPSPDTDVPLSSVEIENPETSDQ lecg_arahy Galactose-binding lectin Agglutinin PNA protein protein arahy Arachis hypogaea (Peanut) MKPFCVFLTFFLLLAASSKKVDSAETVSFNFNSFSEGNPAINFQGDVTVLSNGNIQLTNLNKVNSVGRVLYAMPVRIWSSATGNVASFLTSFSFEMKDIKDYDPADGIIFFIAPEDTQIPAGSIGGGTLGVSDTKGAGHFVGVEFDTYSNSEYNDPPTDHVGIDVNSVDSVKTVPWNSVSGAVVKVTVIYDSSTKTLSVAVTNDNGDITTIAQVVDLKAKLPERVKFGFSASGSLGGRQIHLIRSWSFTSTLITTTRRSIDNNEKKIMNMASA eba1_plafc Erythrocyte-binding antigen 175 protein protein isolate Camp / Malaysia Plasmodium falciparum MKCNISIYFFASFFVLYFAKARNEYDIKENEKFLDVYKEKFNELDKKKYGNVQKTDKKIFTFIENKLDILNNSKFNKRWKSYGTPDNIDKNMSLINKHNNEEMFNNNYQSFLSTSSLIKQNKYVPINAVRVSRILSFLDSRINNGRNTSSNNEVLSNCREKRKGMKWDCKKKNDRSNYVCIPDRRIQLCIVNLSIIKTYTKETMKDHFIEASKKESQLLLKKNDNKYNSKFCNDLKNSFLDYGHLAMGNDMDFGGYSTKAENKIQEVFKGAHGEISEHKIKNFRKEWWNEFREKLWEAMLSEHKNNINNCKNIPQEELQITQWIKEWHGEFLLERDNRSKLPKSKCKNNTLYEACEKECIDPCMKYRDWIIRSKFEWHTLSKEYETQKVPKENAENYLIKISENKNDAKVSLLLNNCDAEYSKYCDCKHTTTLVKSVLNGNDNTIKEKREHIDLDDFSKFGCDKNSVDTNTKVWECKNPYILSTKDVCVPPRRQELCLGNIDRIYDKNLLMIKEHILAIAIYESRILKRKYKNKDDKEVCKIINKTFADIRDIIGGTDYWNDLSNRKLVGKINTNSKYVHRNKKNDKLFRDEWWKVIKKDVWNVISWVFKDKTVCKEDDIENIPQFFRWFSEWGDDYCQDKTKMIETLKVECKEKPCEDDNCKSKCNSYKEWISKKKEEYNKQAKQYQEYQKGNNYKMYSEFKSIKPEVYLKKYSEKCSNLNFEDEFKEELHSDYKNKCTMCPEVKDVPISIIRNNEQTSQEAVPEENTEIAHRTETPSISEGPKGNEQKERDDDSLSKISVSPENSRPETDAKDTSNLLKLKGDVDISMPKAVIGSSPNDNINVTEQGDNISGVNSKPLSDDVRPDKKELEDQNSDESEETVVNHISKSPSINNGDDSGSGSATVSESSSSNTGLSIDDDRNGDTFVRTQDTANTEDVIRKENADKDEDEKGADEERHSTSESLSSPEEKMLTDNEGGNSLNHEEVKEHTSNSDNVQQSGGIVNMNVEKELKDTLENPSSSLDEGKAHEELSEPNLSSDQDMSNTPGPLDNTSEETTERISNNEYKVNEREDERTLTKEYEDIVLKSHMNRESDDGELYDENSDLSTVNDESEDAEAKMKGNDTSEMSHNSSQHIESDQQKNDMKTVGDLGTTHVQNEISVPVTGEIDEKLRESKESKIHKAEEERLSHTDIHKINPEDRNSNTLHLKDIRNEENERHLTNQNINISQERDLQKHGFHTMNNLHGDGVSERSQINHSHHGNRQDRGGNSGNVLNMRSNNNNFNNIPSRYNLYDKKLDLDLYENRNDSTTKELIKKLAEINKCENEISVKYCDHMIHEEIPLKTCTKEKTRNLCCAVSDYCMSYFTYDSEEYYNCTKREFDDPSYTCFRKEAFSSMIFKFLITNKIYYYFYTYKTAKVTIKKINFSLIFFFFFSF gypa-le1-1 3021257 N/A 3021259 unspecified role unspecified role prey prey region alexa 488 label alexa 488 label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false soybn Glycine max (Soybean) Glycine hispida N/A 3021260 unspecified role unspecified role bait bait purified purified region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd_delta_exon3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f physical association physical association false false false Figure 3 C/D SBA from Glycine max binds to alpha- and beta-N-acetyl-galactosamine (GalNAc). galactose-lecg_arahy-1 3021257 N/A 3021261 unspecified role unspecified role prey prey region alexa 488 label alexa 488 label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false arahy Arachis hypogaea (Peanut) N/A 3021260 unspecified role unspecified role bait bait purified purified region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd_delta_exon3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f physical association physical association false false false Figure 3 e/f It is stated that PNA from Arachis hypogaea is specific for beta-galactose (or beta-galactose linked to N-Acetylgalactosamine). gypa-lecg_arahy-1 3021258 N/A 3021262 unspecified role unspecified role prey prey P19214:p.[Asn147Gln;Ser194Ala;Ser339Ala;Thr350Ala;Asn544Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false RII_region sufficient to bind sufficient binding region sufficient to bind range range range range false picpa Pichia pastoris (Yeast) Yeast Pichia pastoris N/A 3021260 unspecified role unspecified role bait bait purified purified P02724:p.Ser66Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P02724:p.Thr69Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region glycores glycosylated residue GlycoRes undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 fc-igg tag fc-igg tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecd_delta_exon3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P02724:p.Ser63Ala 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 P02724:p.Thr69Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P02724:p.[Ser63Ala;Ser66Ala;Thr69Ala] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false exon3 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f physical association physical association false false false Figure 4, Supp fig 3 neuraminidase treatment