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/ The role of the LB structural loop and its interactions with the PDZ domain of the human HtrA3 protease. The role of the LB structural loop and its interactions with the PDZ domain of the human HtrA3 protease. Biochimica et biophysica acta. Proteins and proteomics 2017 imex curation Wenta T., Glaza P., Jarząb M., Zarzecka U., Żurawa-Janicka D., Lesner A., Skórko-Glonek J., Lipińska B. barbara.lipinska@biol.ug.edu.pl 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro protease assay protease assay predetermined predetermined participant predetermined Wenta T., Glaza P., Jarząb M., Zarzecka U., Żurawa-Janicka D., Lesner A., Skórko-Glonek J., Lipińska B. Beta Casein has been assumed to be of Bovine (milk) origin as this protein is used in multiple publications by this group. barbara.lipinska@biol.ug.edu.pl Accepted 2020-APR-02 AT 12:49 BST AT 12:49 BST by LIVIAP Biochimica et biophysica acta. Proteins and proteomics 2017 Only protein-protein interactions imex curation imex curation The role of the LB structural loop and its interactions with the PDZ domain of the human HtrA3 protease. The role of the LB structural loop and its interactions with the PDZ domain of the human HtrA3 protease. Biochimica et biophysica acta. Proteins and proteomics 2017 imex curation Wenta T., Glaza P., Jarząb M., Zarzecka U., Żurawa-Janicka D., Lesner A., Skórko-Glonek J., Lipińska B. barbara.lipinska@biol.ug.edu.pl 04-05-20: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight silver stain molecular weight estimation by silver staining weight silver stain Wenta T., Glaza P., Jarząb M., Zarzecka U., Żurawa-Janicka D., Lesner A., Skórko-Glonek J., Lipińska B. barbara.lipinska@biol.ug.edu.pl Accepted 2020-APR-02 AT 12:49 BST AT 12:49 BST by LIVIAP Biochimica et biophysica acta. Proteins and proteomics 2017 Only protein-protein interactions imex curation imex curation htra1_human Serine protease HTRA1 HTRA1 HTRA PRSS11 High-temperature requirement A serine peptidase 1 L56 Serine protease 11 protein protein human Homo sapiens Human MQIPRAALLPLLLLLLAAPASAQLSRAGRSAPLAAGCPDRCEPARCPPQPEHCEGGRARDACGCCEVCGAPEGAACGLQEGPCGEGLQCVVPFGVPASATVRRRAQAGLCVCASSEPVCGSDANTYANLCQLRAASRRSERLHRPPVIVLQRGACGQGQEDPNSLRHKYNFIADVVEKIAPAVVHIELFRKLPFSKREVPVASGSGFIVSEDGLIVTNAHVVTNKHRVKVELKNGATYEAKIKDVDEKADIALIKIDHQGKLPVLLLGRSSELRPGEFVVAIGSPFSLQNTVTTGIVSTTQRGGKELGLRNSDMDYIQTDAIINYGNSGGPLVNLDGEVIGINTLKVTAGISFAIPSDKIKKFLTESHDRQAKGKAITKKKYIGIRMMSLTSSKAKELKDRHRDFPDVISGAYIIEVIPDTPAEAGGLKENDVIISINGQSVVSANDVSDVIKRESTLNMVVRRGNEDIMITVIPEEIDP DIP protein Q92743 original sequence version: 106 DIP protein Q92743 original sequence version: 149 DIP protein Q92743 original sequence version: 119 CA20A99480FB2330 xiap_human E3 ubiquitin-protein ligase XIAP Inhibitor of apoptosis protein 3 X-linked inhibitor of apoptosis protein IAP-like protein XIAP API3 IAP3 BIRC4 Baculoviral IAP repeat-containing protein 4 RING-type E3 ubiquitin transferase XIAP protein protein human Homo sapiens Human MTFNSFEGSKTCVPADINKEEEFVEEFNRLKTFANFPSGSPVSASTLARAGFLYTGEGDTVRCFSCHAAVDRWQYGDSAVGRHRKVSPNCRFINGFYLENSATQSTNSGIQNGQYKVENYLGSRDHFALDRPSETHADYLLRTGQVVDISDTIYPRNPAMYSEEARLKSFQNWPDYAHLTPRELASAGLYYTGIGDQVQCFCCGGKLKNWEPCDRAWSEHRRHFPNCFFVLGRNLNIRSESDAVSSDRNFPNSTNLPRNPSMADYEARIFTFGTWIYSVNKEQLARAGFYALGEGDKVKCFHCGGGLTDWKPSEDPWEQHAKWYPGCKYLLEQKGQEYINNIHLTHSLEECLVRTTEKTPSLTRRIDDTIFQNPMVQEAIRMGFSFKDIKKIMEEKIQISGSNYKSLEVLVADLVNAQKDSMQDESSQTSLQKEISTEEQLRRLQEEKLCKICMDRNIAIVFVPCGHLVTCKQCAEAVDKCPMCYTVITFKQKIFMS DIP protein P98170 original sequence version: 168 DIP protein P98170 original sequence version: 118 DIP protein P98170 original sequence version: 153 9D394C16D45EB635 casb_bovin Beta-casein CSN2 protein protein bovin Bos taurus (Bovine) Bovine MKVLILACLVALALARELEELNVPGEIVESLSSSEESITRINKKIEKFQSEEQQQTEDELQDKIHPFAQTQSLVYPFPGPIPNSLPQNIPPLTQTPVVVPPFLQPEVMGVSKVKEAMAPKHKEMPFPKYPVEPFTESQSLTLTDVENLHLPLPLLQSWMHQPHQPLPPTVMFPPQSVLSLSQSKVLPVPQKAVPYPQRDMPIQAFLLYQEPVLGPVRGPFPIIV DIP protein P02666 original sequence version: 88 DIP protein P02666 original sequence version: 101 F0BBDD8148A238AE p83110-1 Serine protease HTRA3 HTRA3 PRSP High-temperature requirement factor A3 Pregnancy-related serine protease Long pL protein protein human Homo sapiens Human MQARALLLAALAALALAREPPAAPCPARCDVSRCPSPRCPGGYVPDLCNCCLVCAASEGEPCGGPLDSPCGESLECVRGLCRCRWSHAVCGTDGHTYANVCALQAASRRALQLSGTPVRQLQKGACPLGLHQLSSPRYKFNFIADVVEKIAPAVVHIELFLRHPLFGRNVPLSSGSGFIMSEAGLIITNAHVVSSNSAAPGRQQLKVQLQNGDSYEATIKDIDKKSDIATIKIHPKKKLPVLLLGHSADLRPGEFVVAIGSPFALQNTVTTGIVSTAQREGRELGLRDSDMDYIQTDAIINYGNSGGPLVNLDGEVIGINTLKVTAGISFAIPSDRITRFLTEFQDKQIKDWKKRFIGIRMRTITPSLVDELKASNPDFPEVSSGIYVQEVAPNSPSQRGGIQDGDIIVKVNGRPLVDSSELQEAVLTESPLLLEVRRGNDDLLFSIAPEVVM 3046FCDA1AB24FA6 htra2_human Serine protease HTRA2, mitochondrial High temperature requirement protein A2 Omi stress-regulated endoprotease Serine proteinase OMI Serine protease 25 HTRA2 PRSS25 OMI protein protein human Homo sapiens Human MAAPRAGRGAGWSLRAWRALGGIRWGRRPRLTPDLRALLTSGTSDPRARVTYGTPSLWARLSVGVTEPRACLTSGTPGPRAQLTAVTPDTRTREASENSGTRSRAWLAVALGAGGAVLLLLWGGGRGPPAVLAAVPSPPPASPRSQYNFIADVVEKTAPAVVYIEILDRHPFLGREVPISNGSGFVVAADGLIVTNAHVVADRRRVRVRLLSGDTYEAVVTAVDPVADIATLRIQTKEPLPTLPLGRSADVRQGEFVVAMGSPFALQNTITSGIVSSAQRPARDLGLPQTNVEYIQTDAAIDFGNSGGPLVNLDGEVIGVNTMKVTAGISFAIPSDRLREFLHRGEKKNSSSGISGSQRRYIGVMMLTLSPSILAELQLREPSFPDVQHGVLIHKVILGSPAHRAGLRPGDVILAIGEQMVQNAEDVYEAVRTQSQLAVQIRRGRETLTLYVTPEVTE CEA955A7D0DD8C0D ala(mca)irrvsysf–anb–nh2 peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) IRRVSYSF degp_ecoli Periplasmic serine endoprotease DegP degP htrA ptd b0161 JW0157 Heat shock protein DegP Protease Do protein protein ecoli Escherichia coli (strain K12) MKKTTLALSALALSLGLALSPLSATAAETSSATTAQQMPSLAPMLEKVMPSVVSINVEGSTTVNTPRMPRNFQQFFGDDSPFCQEGSPFQSSPFCQGGQGGNGGGQQQKFMALGSGVIIDADKGYVVTNNHVVDNATVIKVQLSDGRKFDAKMVGKDPRSDIALIQIQNPKNLTAIKMADSDALRVGDYTVAIGNPFGLGETVTSGIVSALGRSGLNAENYENFIQTDAAINRGNSGGALVNLNGELIGINTAILAPDGGNIGIGFAIPSNMVKNLTSQMVEYGQVKRGELGIMGTELNSELAKAMKVDAQRGAFVSQVLPNSSAAKAGIKAGDVITSLNGKPISSFAALRAQVGTMPVGSKLTLGLLRDGKQVNVNLELQQSSQNQVDSSSIFNGIEGAEMSNKGKDQGVVVNNVKTGTPAAQIGLKKGDVIIGANQQAVKNIAELRKVLDSKPSVLALNIQRGDSTIYLLMQ DIP protein P0C0V0 original sequence version: 94 DIP protein P0C0V0 original sequence version: 75 DIP protein P0C0V0 original sequence version: 62 DIP protein P0C0V0 original sequence version: 84 DIP protein P0C0V0 original sequence version: 67 5482E596F74B6D5F htra3-17 2 6 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component 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 P83110-1:p.Phe142Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain F D Tue Jun 30 18:05:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe142Asp' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P83110-1:p.Glu371Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 16:36:15 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu371Ala' P83110-1:p.Gln389Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Q A Tue Jun 30 20:14:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln389Ala' P83110-1:p.Arg362Ala mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 20:03:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg362Ala' physical association physical association 3A/C/D,7A, Table 4. Supp. fig. 1. htra3-18 2 6 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component 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 region_minus_LB sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain P83110-1:p.[Arg362Ala;Glu371Ala;Gln389Ala] mutation mutation certain certain sequence position certain certain certain sequence position certain E A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain Q A Tue Jun 30 15:35:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg362Ala;Glu371Ala;Gln389Ala]' physical association physical association 3A/B, Table 4. Supp. fig. 1 htra3-19 2 3 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Supp. fig. 1, Table 4. htra2-1 2 7 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Supp. fig. 1, Table 4. htra3-20 1 5 enzyme target enzyme target substrate substrate neutral component neutral component purified purified bovin-milk Bos taurus milk milk 6 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified P83110-1:p.Phe142Asp mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain F D Tue Jun 30 19:48:04 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe142Asp' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain p.Asn196_Gly201del mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain NSAAPG . P83110-1:p.[Arg362Ala;Glu371Ala;Gln389Ala] 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 E A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain Q A Tue Jun 30 17:50:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg362Ala;Glu371Ala;Gln389Ala]' 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 P83110-1:p.Ser305Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 17:33:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser305Ala' ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Figure 4, 7B/C, 8,Supp. fig 2. Cleavage rate values for Delta-N-HtrA3L was: 3.7 nM b-casein/uM enzyme/min. htra3-xiap-17 1 4 enzyme target enzyme target substrate substrate neutral component neutral component 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 ecoli Escherichia coli (strain K12) 6 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined enzyme enzyme 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 P83110-1:p.Ser305Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 20:19:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser305Ala' p.Asn196_Gly201del mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain NSAAPG . P83110-1:p.[Arg362Ala;Glu371Ala;Gln389Ala] 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 E A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain Q A Tue Jun 30 16:02:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg362Ala;Glu371Ala;Gln389Ala]' 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Figure 4, 8, Supp. fig 2. Cleavage rate values for Delta-N-HtrA3L was: 60.1nM XIAP/uM enzyme/min. ala(mca)irrvsysfanbnh2-htra3-8 1 8 enzyme target enzyme target substrate substrate neutral component neutral component purified purified N-terminus fluorophore fluorophore n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified P83110-1:p.Arg362Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain R A Tue Jun 30 18:36:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg362Ala' P83110-1:p.Glu371Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain E A Tue Jun 30 17:05:58 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu371Ala' P83110-1:p.Gln389Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain Q A Tue Jun 30 15:54:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln389Ala' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain p.Asn196_Gly201del mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain NSAAPG . P83110-1:p.[Arg362Ala;Glu371Ala;Gln389Ala] mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain E A certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain Q A Tue Jun 30 18:30:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg362Ala;Glu371Ala;Gln389Ala]' 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Table 5, Figure 5, Supp. fig 3 k cat /Km (units /M/sec) (catalytic efficiency) for Wt =38,160. Mutant values range between ~10,000 to ~16, 000. At temperatures 25–40 deg.C all mutations had a negative effect on the peptide cleavage rate but as the temperature increased, the effect decreased. However, the effect of the DeltaLB mutation could not be alleviated. htra3-xiap-18 1 4 enzyme target enzyme target substrate substrate neutral component neutral component 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 ecoli Escherichia coli (strain K12) 3 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined enzyme enzyme 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 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Figure 8 htra1-xiap-1 1 4 enzyme target enzyme target substrate substrate neutral component neutral component 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 ecoli Escherichia coli (strain K12) 7 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined enzyme enzyme 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 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Figure 8 htra1-xiap-2 1 5 enzyme target enzyme target substrate substrate neutral component neutral component purified purified bovin-milk Bos taurus milk milk 3 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined enzyme enzyme 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 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Figure 8 csn2-htra1-1 1 5 enzyme target enzyme target substrate substrate neutral component neutral component purified purified bovin-milk Bos taurus milk milk 7 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined enzyme enzyme 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 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Figure 8 htra2-xiap-6 1 5 enzyme target enzyme target substrate substrate neutral component neutral component purified purified bovin-milk Bos taurus milk milk 9 weight silver stain molecular weight estimation by silver staining weight silver stain predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component 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 ecoli Escherichia coli (strain K12) protein cleavage protein cleavage Figure 8 The bacterial HtrA(DegP) very efficiently digested beta-casein but was practically unable to cut XIAP.