UniProt UniProt http://www.uniprot.org huang-2010-13 in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight by comassie molecular weight estimation by coomasie staining weight by comassie Huang X., Dementiev A., Olson ST., Gettins PG. pgettins@uic.edu The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation huang-2010-14 in vitro In vitro affinity techniques affinity technology affinity techniques predetermined predetermined participant predetermined Huang X., Dementiev A., Olson ST., Gettins PG. Binding of inhibitor to enzyme was allowed sufficient time to reach maximal inhibition, based on measured second order association rate constants. Aliquots of the complex were then diluted into an artificial fluorogenic substrate. Complex dissociation was calculated from increases in absorbance or fluorescence and dissociation rate constants were obtained by fitting progress curves to a parabolic function. pgettins@uic.edu The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation huang-2010-16 in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Huang X., Dementiev A., Olson ST., Gettins PG. Mutated protein tagged with iodoacetamido-N,N′-dimethyl-N-(acetyl)-N′-(7-nitrobenz-3-oxa-1,3-diazol-4-yl)ethylenediamine (NBD). Change in fluorescence on PROZ binding was monitored. pgettins@uic.edu The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation huang-2010-17 in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Huang X., Dementiev A., Olson ST., Gettins PG. Changes in reactivity of mutated protein to iodoacetamido-N,N′-dimethyl-N-(acetyl)-N′-(7-nitrobenz-3-oxa-1,3-diazol-4-yl)ethylenediamine (NBD) on PROZ binding was monitored. pgettins@uic.edu The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation huang-2010-12 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Huang X., Dementiev A., Olson ST., Gettins PG. pgettins@uic.edu The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation huang-2010-15 in vitro In vitro affinity techniques affinity technology affinity techniques predetermined predetermined participant predetermined Huang X., Dementiev A., Olson ST., Gettins PG. Synthetic phospholipid vesicles consisting of a mixture of 70% dioleoylphosphatidylcholine and 30% dioleoylphosphatidylserine were prepared. Binding to vesicle bound-PROZ was measured by changes in scattered light intensity. PROZ binding to the vesicles was Calcium dependent. pgettins@uic.edu The Journal of biological chemistry 2010 Only protein-protein interactions imex curation imex curation proz_human Vitamin K-dependent protein Z PROZ protein protein human Homo sapiens Human MAGCVPLLQGLVLVLALHRVEPSVFLPASKANDVLVRWKRAGSYLLEELFEGNLEKECYEEICVYEEAREVFENEVVTDEFWRRYKGGSPCISQPCLHNGSCQDSIWGYTCTCSPGYEGSNCELAKNECHPERTDGCQHFCLPGQESYTCSCAQGYRLGEDHKQCVPHDQCACGVLTSEKRAPDLQDLPWQVKLTNSEGKDFCGGVIIRENFVLTTAKCSLLHRNITVKTYFNRTSQDPLMIKITHVHVHMRYDADAGENDLSLLELEWPIQCPGAGLPVCTPEKDFAEHLLIPRTRGLLSGWARNGTDLGNSLTTRPVTLVEGEECGQVLNVTVTTRTYCERSSVAAMHWMDGSVVTREHRGSWFLTGVLGSQPVGGQAHMVLVTKVSRYSLWFKQIMN zpi_human Protein Z-dependent protease inhibitor SERPINA10 ZPI Serpin A10 UNQ707/PRO1358 protein protein human Homo sapiens Human MKVVPSLLLSVLLAQVWLVPGLAPSPQSPETPAPQNQTSRVVQAPKEEEEDEQEASEEKASEEEKAWLMASRQQLAKETSNFGFSLLRKISMRHDGNMVFSPFGMSLAMTGLMLGATGPTETQIKRGLHLQALKPTKPGLLPSLFKGLRETLSRNLELGLTQGSFAFIHKDFDVKETFFNLSKRYFDTECVPMNFRNASQAKRLMNHYINKETRGKIPKLFDEINPETKLILVDYILFKGKWLTPFDPVFTEVDTFHLDKYKTIKVPMMYGAGKFASTFDKNFRCHVLKLPYQGNATMLVVLMEKMGDHLALEDYLTTDLVETWLRNMKTRNMEVFFPKFKLDQKYEMHELLRQMGIRRIFSPFADLSELSATGRNLQVSRVLQRTVIEVDERGTEAVAGILSEITAYSMPPVIKVDRPFHFMIYEETSGMLLFLGRVVNPTLL fa10_human Coagulation factor X Stuart factor Stuart-Prower factor F10 protein protein human Homo sapiens Human MGRPLHLVLLSASLAGLLLLGESLFIRREQANNILARVTRANSFLEEMKKGHLERECMEETCSYEEAREVFEDSDKTNEFWNKYKDGDQCETSPCQNQGKCKDGLGEYTCTCLEGFEGKNCELFTRKLCSLDNGDCDQFCHEEQNSVVCSCARGYTLADNGKACIPTGPYPCGKQTLERRKRSVAQATSSSGEAPDSITWKPYDAADLDPTENPFDLLDFNQTQPERGDNNLTRIVGGQECKDGECPWQALLINEENEGFCGGTILSEFYILTAAHCLYQAKRFKVRVGDRNTEQEEGGEAVHEVEVVIKHNRFTKETYDFDIAVLRLKTPITFRMNVAPACLPERDWAESTLMTQKTGIVSGFGRTHEKGRQSTRLKMLEVPYVDRNSCKLSSSFIITQNMFCAGYDTKQEDACQGDSGGPHVTRFKDTYFVTGIVSWGEGCARKGKYGIYTKVTAFLKWIDRSMKTRGLPKAKSHAPEVITSSPLK p03951-pro_0000027826 Coagulation factor XIa light chain F11 Plasma thromboplastin antecedent protein protein human Homo sapiens Human IVGGTASVRGEWPWQVTLHTTSPTQRHLCGGSIIGNQWILTAAHCFYGVESPKILRVYSGILNQSEIKEDTSFFGVQEIIIHDQYKMAESGYDIALLKLETTVNYTDSQRPICLPSKGDRNVIYTDCWVTGWGYRKLRDKIQNTLQKAKIPLVTNEECQKRYRGHKITHKMICAGYREGGKDACKGDSGGPLSCKHNEVWHLVGITSWGEGCAQRERPGVYTNVVEYVDWILEKTQAV 388 625 p03951-pro_0000027825 Coagulation factor XIa heavy chain F11 Plasma thromboplastin antecedent protein protein human Homo sapiens Human ECVTQLLKDTCFEGGDITTVFTPSAKYCQVVCTYHPRCLLFTFTAESPSEDPTRWFTCVLKDSVTETLPRVNRTAAISGYSFKQCSHQISACNKDIYVDLDMKGINYNSSVAKSAQECQERCTDDVHCHFFTYATRQFPSLEHRNICLLKHTQTGTPTRITKLDKVVSGFSLKSCALSNLACIRDIFPNTVFADSNIDSVMAPDAFVCGRICTHHPGCLFFTFFSQEWPKESQRNLCLLKTSESGLPSTRIKKSKALSGFSLQSCRHSIPVFCHSSFYHDTDFLGEELDIVAAKSHEACQKLCTNAVRCQFFTYTPAQASCNEGKGKCYLKLSSNGSPTKILHGRGGISGYTLRLCKMDNECTTKIKPR 19 387 proz-serpina10-2 3635918 N/A 3635924 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-plasma homo sapiens blood plasma plasma blood plasma N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified Q9UK55:p.[Asp95Ala;Asp259_Lys260delinsAlaAla;Glu404Ala] 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.[Glu334Ala;Glu404Ala] 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 certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.Glu334Ala 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 Q9UK55:p.Glu404Ala 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 spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction false false false Fig. 2 f10-serpina10-2 3635918 N/A 3635926 unspecified role unspecified role neutral component neutral component purified purified N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction false false false Fig. 8 f10-serpina10-1 3635919 N/A 3635926 unspecified role unspecified role neutral component neutral component purified purified N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified Q9UK55:p.Glu334Ala 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 Q9UK55:p.Glu404Ala 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 Q9UK55:p.[Glu334Ala;Glu404Ala] 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 certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.[Asp95Ala;Asp259_Lys260delinsAlaAla;Asp314Ala] 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 certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.Tyr408Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false Table 1-3, Figs. 5, 8 In the presence of PROZ, lipid, and Ca2+ cofactors, there was a 10-fold reduction in F10-SERPINA10 binding for the F10 p.Glu334Ala mutant but the Glu404Ala mutant still had no effect. f11-serpina10-1 3635919 N/A 3635927 unspecified role unspecified role unspecified role unspecified role purified purified heavy chain binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3635928 unspecified role unspecified role neutral component neutral component purified purified light chain binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified Q9UK55:p.Tyr408Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false 3635953 3635951 physical association physical association false false false 3635919 Table 2 Carried out in the absence of cofactors. proz-serpina10-4 3635920 N/A 3635924 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-plasma homo sapiens blood plasma plasma blood plasma Stoichiometry: 1.0 N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.Leu402Cys mutation mutation certain certain sequence position certain certain certain sequence position certain false fluorescence tagged residue fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm Stoichiometry: 1.0 direct interaction direct interaction false false false 3635920 Fig. 6A proz-serpina10-5 3635920 N/A 3635924 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-plasma homo sapiens blood plasma plasma blood plasma Stoichiometry: 1.0 N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.Ser143Cys mutation mutation certain certain sequence position certain certain certain sequence position certain false fluorescence tagged residue fluorescent dye fluorescent dye label fluorescent dye certain certain sequence position certain certain certain sequence position certain false spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm Stoichiometry: 1.0 direct interaction direct interaction false false false 3635920 Fig. 6B proz-serpina10-6 3635921 N/A 3635924 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-plasma homo sapiens blood plasma plasma blood plasma N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.Leu402Cys mutation mutation certain certain sequence position certain certain certain sequence position certain false spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction false false false Fig. 7 proz-serpina10-7 3635921 N/A 3635924 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-plasma homo sapiens blood plasma plasma blood plasma N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9UK55:p.Ser143Cys mutation mutation certain certain sequence position certain certain certain sequence position certain false spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction false false false Fig. 7 proz-serpina10-1 3635922 N/A 3635924 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-plasma homo sapiens blood plasma plasma blood plasma Stoichiometry: 1.0 N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain 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 Stoichiometry: 1.0 direct interaction direct interaction false false false Supp. Table 1 proz-serpina10-3 3635923 N/A 3635924 unspecified role unspecified role neutral component neutral component purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-plasma homo sapiens blood plasma plasma blood plasma N/A 3635925 unspecified role unspecified role neutral component neutral component purified purified Q9UK55:p.[Asp95Ala;Asp259_Lys260delinsAlaAla;Glu404Ala] 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false spofr Spodoptera frugiperda (Fall armyworm) Fall armyworm direct interaction direct interaction false false false Fig. 3