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/ poulsen-2019-1 in vitro In vitro protease assay protease assay experimental particp experimental participant identification experimental particp Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ., Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ. Rare diseases - Interactions investigated in the context of Rare genetic disease jje@mbg.au.dk The Journal of biological chemistry 2019 Only protein-protein interactions imex curation imex curation poulsen-2019-2 in vitro In vitro aggregation aggregation assay aggregation sequence tag sequence tag identification sequence tag Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ., Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ. Rare diseases - Interactions investigated in the context of Rare genetic disease Mutant samples were incubated with HtrA1 for 21 hours. After an additional 4 days of sample incubation, to allow the degradation products to form fibrils, the pellets were separated from the supernatant by centrifugation. jje@mbg.au.dk The Journal of biological chemistry 2019 Only protein-protein interactions imex curation imex curation poulsen-2019-3 in vitro In vitro aggregation aggregation assay aggregation predetermined predetermined participant predetermined Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ., Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ. Rare diseases - Interactions investigated in the context of Rare genetic disease FAS1-4 protein was incubated with and without the presence of HtrA1. Fibril formation was monitored using a continuous thioflavin T assay. When ThT binds to beta sheet-rich structures, such as those in amyloid aggregates, the dye displays enhanced fluorescence and a characteristic red shift of its emission spectrum. Binding induces a shift in excitation maximum (385 nm to 450 nm) and emission maximum (445 nm to 482 nm). jje@mbg.au.dk The Journal of biological chemistry 2019 Only protein-protein interactions imex curation imex curation poulsen-2019-4 in vitro In vitro ir spectrometry infrared spectroscopy ir spectrometry predetermined predetermined participant predetermined Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ., Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ. Rare diseases - Interactions investigated in the context of Rare genetic disease Fourier transform infrared (FTIR) spectroscopy jje@mbg.au.dk The Journal of biological chemistry 2019 Only protein-protein interactions imex curation imex curation poulsen-2019-5 in vitro In vitro tem transmission electron microscopy tem predetermined predetermined participant predetermined Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ., Poulsen ET., Nielsen NS., Scavenius C., Mogensen EH., Risør MW., Runager K., Lukassen MV., Rasmussen CB., Christiansen G., Richner M., Vorum H., Enghild JJ. Rare diseases - Interactions investigated in the context of Rare genetic disease jje@mbg.au.dk The Journal of biological chemistry 2019 Only protein-protein interactions imex curation imex curation q92743-pro_0000026943 Serine protease HTRA1 HTRA1 HTRA PRSS11 High-temperature requirement A serine peptidase 1 L56 Serine protease 11 protein protein human Homo sapiens Human QLSRAGRSAPLAAGCPDRCEPARCPPQPEHCEGGRARDACGCCEVCGAPEGAACGLQEGPCGEGLQCVVPFGVPASATVRRRAQAGLCVCASSEPVCGSDANTYANLCQLRAASRRSERLHRPPVIVLQRGACGQGQEDPNSLRHKYNFIADVVEKIAPAVVHIELFRKLPFSKREVPVASGSGFIVSEDGLIVTNAHVVTNKHRVKVELKNGATYEAKIKDVDEKADIALIKIDHQGKLPVLLLGRSSELRPGEFVVAIGSPFSLQNTVTTGIVSTTQRGGKELGLRNSDMDYIQTDAIINYGNSGGPLVNLDGEVIGINTLKVTAGISFAIPSDKIKKFLTESHDRQAKGKAITKKKYIGIRMMSLTSSKAKELKDRHRDFPDVISGAYIIEVIPDTPAEAGGLKENDVIISINGQSVVSANDVSDVIKRESTLNMVVRRGNEDIMITVIPEEIDP 23 480 q15582-pro_0000008769 Transforming growth factor-beta-induced protein ig-h3 TGFBI BIGH3 Kerato-epithelin RGD-containing collagen-associated protein protein protein human Homo sapiens Human GPAKSPYQLVLQHSRLRGRQHGPNVCAVQKVIGTNRKYFTNCKQWYQRKICGKSTVISYECCPGYEKVPGEKGCPAALPLSNLYETLGVVGSTTTQLYTDRTEKLRPEMEGPGSFTIFAPSNEAWASLPAEVLDSLVSNVNIELLNALRYHMVGRRVLTDELKHGMTLTSMYQNSNIQIHHYPNGIVTVNCARLLKADHHATNGVVHLIDKVISTITNNIQQIIEIEDTFETLRAAVAASGLNTMLEGNGQYTLLAPTNEAFEKIPSETLNRILGDPEALRDLLNNHILKSAMCAEAIVAGLSVETLEGTTLEVGCSGDMLTINGKAIISNKDILATNGVIHYIDELLIPDSAKTLFELAAESDVSTAIDLFRQAGLGNHLSGSERLTLLAPLNSVFKDGTPPIDAHTRNLLRNHIIKDQLASKYLYHGQTLETLGGKKLRVFVYRNSLCIENSCIAAHDKRGRYGTLFTMDRVLTPPMGTVMDVLKGDNRFSMLVAAIQSAGLTETLNREGVYTVFAPTNEAFRALPPRERSRLLGDAKELANILKYHIGDEILVSGGIGALVRLKSLQGDKLEVSLKNNVVSVNKEPVAEPDIMATNGVVHVITNVLQPPANRPQERGDELADSALEIFKQASAFSRASQRSVRLAPVYQKLLERMKH 24 683 bgh3_human Transforming growth factor-beta-induced protein ig-h3 TGFBI BIGH3 Kerato-epithelin RGD-containing collagen-associated protein protein protein human Homo sapiens Human MALFVRLLALALALALGPAATLAGPAKSPYQLVLQHSRLRGRQHGPNVCAVQKVIGTNRKYFTNCKQWYQRKICGKSTVISYECCPGYEKVPGEKGCPAALPLSNLYETLGVVGSTTTQLYTDRTEKLRPEMEGPGSFTIFAPSNEAWASLPAEVLDSLVSNVNIELLNALRYHMVGRRVLTDELKHGMTLTSMYQNSNIQIHHYPNGIVTVNCARLLKADHHATNGVVHLIDKVISTITNNIQQIIEIEDTFETLRAAVAASGLNTMLEGNGQYTLLAPTNEAFEKIPSETLNRILGDPEALRDLLNNHILKSAMCAEAIVAGLSVETLEGTTLEVGCSGDMLTINGKAIISNKDILATNGVIHYIDELLIPDSAKTLFELAAESDVSTAIDLFRQAGLGNHLSGSERLTLLAPLNSVFKDGTPPIDAHTRNLLRNHIIKDQLASKYLYHGQTLETLGGKKLRVFVYRNSLCIENSCIAAHDKRGRYGTLFTMDRVLTPPMGTVMDVLKGDNRFSMLVAAIQSAGLTETLNREGVYTVFAPTNEAFRALPPRERSRLLGDAKELANILKYHIGDEILVSGGIGALVRLKSLQGDKLEVSLKNNVVSVNKEPVAEPDIMATNGVVHVITNVLQPPANRPQERGDELADSALEIFKQASAFSRASQRSVRLAPVYQKLLERMKH htra1-tgfbi-1 3671702 N/A 3671707 predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f N/A 3671708 ms participant Identification by mass spectrometry ms participant western blot western blot Immuno blot Immuno blot enzyme target enzyme target substrate substrate 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 Q15582-PRO_0000008769:p.Arg101His 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 Q15582-PRO_0000008769:p.Arg532Trp 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 Q15582-PRO_0000008769:p.[Ala523Asp;Pro528Gln] 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 Q15582-PRO_0000008769:p.Arg101Cys mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false Q15582-PRO_0000008769:p.Ala523Asp mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false Q15582-PRO_0000008769:p.Ala523Thr mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false Q15582-PRO_0000008769:p.Arg101Leu mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f protein cleavage protein cleavage false false false 1B/C/D htra1-tgfbi-2 3671702 N/A 3671707 predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified human-293f Homo sapiens HEK293F cell line 293f embryonic kidney cell 293f N/A 3671709 sequence tag sequence tag identification sequence tag western blot western blot Immuno blot Immuno blot enzyme target enzyme target substrate substrate neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.Arg555Trp 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 Q15582:p.[Ala546Asp;Pro551Gln] mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Arg555Gln 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 Q15582:p.Ala546Thr mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) protein cleavage protein cleavage false false false 2,3 MS mapping of HtrA1 cleavage sites showed that the mutants underwent extensive cleavage in several regions. htra1-tgfbi-3 3671703 N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified amyloidogenic region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified amyloidogenic region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3671743 3671734 3671744 3671735 3671745 3671736 direct interaction direct interaction false false false 4 Peptide content >~10% in pellet. tgfbi-tgfbi-1 3671703 N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 4 Interactions in supernatant. Some supernatant fibril formation was seen for all mutants tested, but not to such a great extent as seen in the pellets. The highest peptide counts were seen outside the Gly-560–Leu-589 region. tgfbi-tgfbi-2 3671704 N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3671775 3671768 direct interaction direct interaction false false false 5A Increased ThT fluorescence as a function of time was observed for all four LCD-linked FAS1-4 variants in the presence of HtrA1. HtrA1(Q92743-PRO_0000026943) tgfbi-tgfbi-4 3671705 N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3671790 3671783 direct interaction direct interaction false false false 5B A shift for the undigested FAS1-4 mutants from 1651-1653 cm-1 to 1627-1630 cm-1, was shown after incubation with HtrA1;values characteristic of beta-sheet and beta-amyloid-sheet formation. HtrA1(Q92743-PRO_0000026943) tgfbi-tgfbi-3 3671706 N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3671709 western blot western blot Immuno blot Immuno blot unspecified role unspecified role neutral component neutral component purified purified FAS 1-4 domains sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15582:p.[Ala546Asp;Pro551Gln] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false Q15582:p.Ala546Thr mutation mutation certain certain sequence position certain certain certain sequence position certain false N-terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q15582:p.Ala546Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false Q15582:p.Val624Met mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 5C Elongated fibrils for FAS1-4 samples incubated with HtrA1were seen.Nonfibrillar aggregates were observed for nonprotease-treated FAS1-4 mutants.