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/ Subcellular discharge of a serine protease mediates release of invasive malaria parasites from host erythrocytes. Subcellular discharge of a serine protease mediates release of invasive malaria parasites from host erythrocytes. Cell (0092-8674) 2007 imex curation Yeoh S., O'Donnell RA., Koussis K., Dluzewski AR., Ansell KH., Osborne SA., Hackett F., Withers-Martinez C., Mitchell GH., Bannister LH., Bryans JS., Kettleborough CA., Blackman MJ. mblackm@nimr.mrc.ac.uk 01-Feb-2008: Contacted by JYOTI. davet Only protein-protein interactions 8 in vitro In vitro protease assay protease assay protein sequence protein sequence identification protein sequence Yeoh S., O'Donnell RA., Koussis K., Dluzewski AR., Ansell KH., Osborne SA., Hackett F., Withers-Martinez C., Mitchell GH., Bannister LH., Bryans JS., Kettleborough CA., Blackman MJ. mblackm@nimr.mrc.ac.uk Accepted 2008-FEB-10 by DAVET. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 8 imex curation Subcellular discharge of a serine protease mediates release of invasive malaria parasites from host erythrocytes. Subcellular discharge of a serine protease mediates release of invasive malaria parasites from host erythrocytes. Cell (0092-8674) 2007 imex curation Yeoh S., O'Donnell RA., Koussis K., Dluzewski AR., Ansell KH., Osborne SA., Hackett F., Withers-Martinez C., Mitchell GH., Bannister LH., Bryans JS., Kettleborough CA., Blackman MJ. mblackm@nimr.mrc.ac.uk 01-Feb-2008: Contacted by JYOTI. davet Only protein-protein interactions 8 in vitro In vitro protease assay protease assay western blot western blot Immuno blot Immuno blot Yeoh S., O'Donnell RA., Koussis K., Dluzewski AR., Ansell KH., Osborne SA., Hackett F., Withers-Martinez C., Mitchell GH., Bannister LH., Bryans JS., Kettleborough CA., Blackman MJ. mblackm@nimr.mrc.ac.uk Accepted 2008-FEB-10 by DAVET. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 8 imex curation Subcellular discharge of a serine protease mediates release of invasive malaria parasites from host erythrocytes. Subcellular discharge of a serine protease mediates release of invasive malaria parasites from host erythrocytes. Cell (0092-8674) 2007 imex curation Yeoh S., O'Donnell RA., Koussis K., Dluzewski AR., Ansell KH., Osborne SA., Hackett F., Withers-Martinez C., Mitchell GH., Bannister LH., Bryans JS., Kettleborough CA., Blackman MJ. mblackm@nimr.mrc.ac.uk 01-Feb-2008: Contacted by JYOTI. davet Only protein-protein interactions 8 in vitro In vitro protease assay protease assay fingerprinting peptide massfingerprinting fingerprinting Yeoh S., O'Donnell RA., Koussis K., Dluzewski AR., Ansell KH., Osborne SA., Hackett F., Withers-Martinez C., Mitchell GH., Bannister LH., Bryans JS., Kettleborough CA., Blackman MJ. Substrates tracked by fluorescently labelling. mblackm@nimr.mrc.ac.uk Accepted 2008-FEB-10 by DAVET. 01-Feb-2008: Contacted by JYOTI. Cell (0092-8674) 2007 davet Only protein-protein interactions 8 imex curation sera5_plaf7 Serine-repeat antigen protein 5 SERA5 111 kDa antigen Serine protease SERA5 p126 PFB0340c protein protein plaf7 Plasmodium falciparum (isolate 3D7) MKSYISLFFILCVIFNKNVIKCTGESQTGNTGGGQAGNTGGDQAGSTGGSPQGSTGASPQGSTGASPQGSTGASQPGSSEPSNPVSSGHSVSTVSVSQTSTSSEKQDTIQVKSALLKDYMGLKVTGPCNENFIMFLVPHIYIDVDTEDTNIELRTTLKKTNNAISFESNSGSLEKKKYVKLPSNGTTGEQGSSTGTVRGDTEPISDSSSSSSSSSSSSSSSSSSSSSSSSSSSESLPANGPDSPTVKPPRNLQNICETGKNFKLVVYIKENTLILKWKVYGETKDTTENNKVDVRKYLINEKETPFTNILIHAYKEHNGTNLIESKNYAIGSDIPEKCDTLASNCFLSGNFNIEKCFQCALLVEKENKNDVCYKYLSEDIVSKFKEIKAETEDDDEDDYTEYKLTESIDNILVKMFKTNENNDKSELIKLEEVDDSLKLELMNYCSLLKDVDTTGTLDNYGMGNEMDIFNNLKRLLIYHSEENINTLKNKFRNAAVCLKNVDDWIVNKRGLVLPELNYDLEYFNEHLYNDKNSPEDKDNKGKGVVHVDTTLEKEDTLSYDNSDNMFCNKEYCNRLKDENNCISNLQVEDQGNCDTSWIFASKYHLETIRCMKGYEPTKISALYVANCYKGEHKDRCDEGSSPMEFLQIIEDYGFLPAESNYPYNYVKVGEQCPKVEDHWMNLWDNGKILHNKNEPNSLDGKGYTAYESERFHDNMDAFVKIIKTEVMNKGSVIAYIKAENVMGYEFSGKKVQNLCGDDTADHAVNIVGYGNYVNSEGEKKSYWIVRNSWGPYWGDEGYFKVDMYGPTHCHFNFIHSVVIFNVDLPMNNKTTKKESKIYDYYLKASPEFYHNLYFKNFNVGKKNLFSEKEDNENNKKLGNNYIIFGQDTAGSGQSGKESNTALESAGTSNEVSERVHVYHILKHIKDGKIRMGMRKYIDTQDVNKKHSCTRSYAFNPENYEKCVNLCNVNWKTCEEKTSPGLCLSKLDTNNECYFCYV CF26A8A03E512ED5 o96164_plaf7 PF3D7_0207700 protein protein plaf7 Plasmodium falciparum (isolate 3D7) MKIHIFLIATIYVLFSEKLIKWTTASTTQGGDTDTHPGTPPGEGSDVSQGAGQDASQGAGQDANPDPTLPKPPSPPADDTKDTGSQGDADSSSSKIEIPPLVKPENHKTIVSAMLKNYKGVKVTGTCGADFGLFLVPHIYVHVKSEDTEIELSSELAPPEMQTKFDKTQLKKFCVKDDTKKFDFIAYIYKDILVFKWKVYEEGLSKEQDVDEMKYLLPNLKQPITSIQVHSWTGTKESYILESKDYVLGEGMPEKCDAIATDCFLSGFTDIGKCFQCKLLMQEKNINDSCFKYVSSNQKELIKKQLKITAQDDEESSEYHLSESIKNLLKNIYKKNNDDNKKKELLHFENVNSALKSELLNYCNLLKEVNMNGVLKDHQLGNVQDVFNNLTKLLEEHKEENDNVLYHKMKNEALCLKNVNDWMKNKTGLLLPQLSYDLTYKNNNFTEFTQNKSYTSQNIVDKLYCNHEYCNRLKDHNNCISKINVEDQKNCALSWAFASKYHLETIKCMKGYEPLNASVLYVTNCLKNKNKDVCTEGSNPLVFLETIEEKGFLPTESNYPYDQSKVGDICPQLQNDWDNVFENTKVLDYNNGPFSVGTKGYIAYESEAFQKDMHSFVKLVKDEIMNKGSVIAYVKAENVLGYELNGKKVQNLCGDKTPDHVVNIVGYGNYINNKGEKKSYWIVRNSWGKYWGDDGYFKVDMYGPSTCEDNFIHTVVVFNVQVPINEKFDKKEHDIYNYYLKTSPEFYHNLYYKTFNSNKEEKSMNKNSYVYGQDTTPVENEAPRSGVQKPTELSSTESQTVSPPNESQTESLLSGGSQVTNPTLTQSTSSSSGQQETGPLSTQGLSPATGDPKGKEQEASPAEGLSGVLNPTKEVTSEEKIQIIHLLKHIKNSKIRRGLVKYNHEFEVGDNSCSRSTSKNAEMHDECVNICEKYWPECRGTAVPGYCLSTHDDKNECDFCYV BF7659CC46D39495 sub1_plaf7 Subtilisin-like protease 1 SUB1 PfSUB1 PF3D7_0507500 protein protein plaf7 Plasmodium falciparum (isolate 3D7) MMLNKKVVALCTLTLHLFCIFLCLGKEVRSEENGKIQDDAKKIVSELRFLEKVEDVIEKSNIGGNEVDADENSFNPDTEVPIEEIEEIKMRELKDVKEEKNKNDNHNNNNNNISSSSSSSSNTFGEEKEEVSKKKKKLRLIVSENHATTPSFFQESLLEPDVLSFLESKGNLSNLKNINSMIIELKEDTTDDELISYIKILEEKGALIESDKLVSADNIDISGIKDAIRRGEENIDVNDYKSMLEVENDAEDYDKMFGMFNESHAATSKRKRHSTNERGYDTFSSPSYKTYSKSDYLYDDDNNNNNYYYSHSSNGHNSSSRNSSSSRSRPGKYHFNDEFRNLQWGLDLSRLDETQELINEHQVMSTRICVIDSGIDYNHPDLKDNIELNLKELHGRKGFDDDNNGIVDDIYGANFVNNSGNPMDDNYHGTHVSGIISAIGNNNIGVVGVDVNSKLIICKALDEHKLGRLGDMFKCLDYCISRNAHMINGSFSFDEYSGIFNSSVEYLQRKGILFFVSASNCSHPKSSTPDIRKCDLSINAKYPPILSTVYDNVISVANLKKNDNNNHYSLSINSFYSNKYCQLAAPGTNIYSTAPHNSYRKLNGTSMAAPHVAAIASLIFSINPDLSYKKVIQILKDSIVYLPSLKNMVAWAGYADINKAVNLAIKSKKTYINSNISNKWKKKSRYLH FACFEFB495AEF4F7 sera6_plaf7 Serine-repeat antigen protein 6 SERA6 Cysteine protease SERA6 PF3D7_0207500 protein protein plaf7 Plasmodium falciparum (isolate 3D7) MICPIFFLYIINVLFTQYFIKCEGNKVTVISHNNGHNDNLDVNKNGVISQENVFDTSESLNLPSNKKVGSDDLNTTTISFTVPDNLENEVKVVSSSESGKGATVSHTKVTSEGLSDTQPNVTQSVSSSTHTPGSLDSTMSTEQHSSVSQSSLPTESSSETLNKATVPEIPIQINSGLLKNYNGVKVTGSCGSYFRVYLVPHILIYALTKYSVIQLESLFNDNARIDVEHKGELQNKCSEGYHFKLVVYITHNVLNLKWKTYKPNEESKSEDSDVRKYRIPKLERPFTSIQVYTANSKAGVIETKNYNIRTDIPDTCDAIATDCFLNGNVNIEKCFQCTLLVQKKDKSHECFKYVSSEMKKKMNEIKVKAQDDFNPNEYKLIESIDNILSKIYKKANKPFEISKDLINLEDLDYQFKNELLEYCKLLKKVDTSGTLEEYELGNAEDIYNNLTRLLKSHSDENIVTLQGKLRNTAICIKNVDEWILNKRGLTLPSESPSESSSKSDSYLNTFNDKDKNEDKDDMSKNSKEEFKNDDKENSDDQNNNDSNKKDDENNINNGDTNYVYDFDDDDYDNNSYEKDMYESPIKENKNGVIDLEKYGNQIKLKSPYFKNSKYCNYEYCNRWRDKTSCISQIEVEEQGNCGLCWIFASKLHFETIRCMRGYGHFRSSALYVANCSKRKPIDRCEEGSNPLEFLRILDEKKFLPLESNYPYSYTSAGNSCPKLPNSWTNLWGDTKLLFNKKVHRYIGNKGFISHETSYFKNNMDLFIDMVKREVQNKGSVIIYIKTQDVIGYDFNGKGVHSMCGDRTPDHAANIIGYGNYINKKGEKRSYWLIRNSWSYYWGDEGNFRVDMLGPKNCLYNFIHTVVFFKLDLGTIHVPKKKSWKKNVYFLRHNPDFMYSLYYNNYEPETSQDFESENDYDNAFVHGQSNESDETNKEGKNVHNSVEKKIQILHILKHIKDSQIKRGLVKYDNINETKDEHTCSRVNSQDAEKYEECKKFCLTKWNECKDHYSPGYCLTDLYKGEDCNFCYV 6DF255195338D316 sub1-sera-1 2 6 enzyme enzyme neutral component neutral component 4 enzyme target enzyme target substrate substrate neutral component neutral component plaf7 Plasmodium falciparum (isolate 3D7) protein cleavage protein cleavage To test directly whether SERA5 can be correctly processed by PfSUB1, SERA5 P126 was purified from schizont extracts (Figure S4) and incubated with recombinant PfSUB1 (rPfSUB1). Fig. 5A and 5B Cleavage of Sera to a P56 form via a P73 intermediate Inhibited by MRT12113, a compound originally purified natural products from temperate plants. sub1-sera-2 1 6 enzyme enzyme neutral component neutral component 4 enzyme target enzyme target substrate substrate neutral component neutral component plaf7 Plasmodium falciparum (isolate 3D7) protein cleavage protein cleavage To test directly whether SERA5 can be correctly processed by PfSUB1, SERA5 P126 was purified from schizont extracts (Figure S4) and incubated with recombinant PfSUB1 (rPfSUB1). Fig. 5C Cleavage of Sera to a P56 form via a P73 intermediate sub1-sera-3 3 4 enzyme target enzyme target substrate substrate neutral component neutral component region fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9TY95:p.Glu386Cys mutation mutation certain certain sequence position certain certain certain sequence position certain E C Wed May 02 18:57:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu386cys' Tue Jun 30 16:55:22 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu386Cys' Q9TY95:p.Asp395Cys mutation mutation certain certain sequence position certain certain certain sequence position certain D C Wed May 02 16:13:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp395cys' Tue Jun 30 19:11:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp395Cys' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9TY95:p.Ile882Cys mutation mutation certain certain sequence position certain certain certain sequence position certain I C Wed May 02 15:46:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile882cys' Tue Jun 30 17:21:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile882Cys' Q9TY95:p.Ser891Cys mutation mutation certain certain sequence position certain certain certain sequence position certain S C Wed May 02 19:45:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser891cys' Tue Jun 30 17:32:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser891Cys' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 6 enzyme enzyme neutral component neutral component protein cleavage protein cleavage To explore this, peptides incorporating SERA5 processing sites 1, 2, and 3 were assessed for susceptibility to PfSUB1 cleavage. Fig 5D and Supp. Fig. 5A sub1 3 6 enzyme enzyme neutral component neutral component 6 enzyme target enzyme target substrate substrate neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q8I0V0:p.Lys212Cys mutation mutation certain certain sequence position certain certain certain sequence position certain K C Wed May 02 14:32:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys212cys' Tue Jun 30 15:37:35 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys212Cys' undefined mutation mutation mutation certain certain sequence position certain certain certain sequence position certain Wed Jul 06 17:46:33 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'region' region fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) protein cleavage protein cleavage They were then compared in kinetic assays with pepF1-6R (6-IATR-labeled Ac-CLVSADNIDIC), a substrate based on the PfSUB1 autocatalytic processing site (Blackman et al., 2002). Fig 5D and Supp. Fig. 5A sub1-sera4-1 2 6 enzyme enzyme neutral component neutral component 5 enzyme target enzyme target substrate substrate neutral component neutral component plaf7 Plasmodium falciparum (isolate 3D7) protein cleavage protein cleavage To test this, full-length SERA4 and SERA6 were isolated from schizont extracts (Figure S4) then incubated with rPfSUB1 as described above for SERA5. Fig. 6B Cleavage of O96164 sub1-sera6-1 2 7 enzyme target enzyme target substrate substrate neutral component neutral component plaf7 Plasmodium falciparum (isolate 3D7) 6 enzyme enzyme neutral component neutral component protein cleavage protein cleavage To test this, full-length SERA4 and SERA6 were isolated from schizont extracts (Figure S4) then incubated with rPfSUB1 as described above for SERA5. Fig. 6B Cleavage of Q9TY96 sub1-sera6-2 3 7 enzyme target enzyme target substrate substrate neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 6 enzyme enzyme neutral component neutral component protein cleavage protein cleavage Synthetic peptides based on predicted SERA4 and SERA6 cleavage sites are correctly cleaved by rPfSUB1. Supp. Fig. 7 Cleavage of Q9TY96 sub1-sera4-2 3 6 enzyme enzyme neutral component neutral component 5 enzyme target enzyme target substrate substrate neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined plaf7 Plasmodium falciparum (isolate 3D7) protein cleavage protein cleavage Synthetic peptides based on predicted SERA4 and SERA6 cleavage sites are correctly cleaved by rPfSUB1. Supp. Fig. 7 Cleavage of O96164