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/ todorova-2017-1 in vitro In vitro aggregation aggregation assay aggregation predetermined predetermined participant predetermined Todorova N., Zlatic CO., Mao Y., Yarovsky I., Howlett GJ., Gooley PR., Griffin MD. 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). Human apolipoprotein C-II was found to have a Uniprot identifier of PRO_0000002024 by mutation mapping. mgriffin@unimelb.edu.au Biochemistry 2017 Only protein-protein interactions imex curation imex curation todorova-2017-3 in vitro In vitro aut-page aut-page predetermined predetermined participant predetermined Todorova N., Zlatic CO., Mao Y., Yarovsky I., Howlett GJ., Gooley PR., Griffin MD. Samples were mixed and incubated under fibril forming conditions, centrifuged, and the supernatant and pellet fractions for each sample were loaded onto an acetic acid-urea PAGE gel for analysis; this method separates apoC-II derivatives based on their net charge at pH 3. Human apolipoprotein C-II was found to have a Uniprot identifier of PRO_0000002024 by mutation mapping. mgriffin@unimelb.edu.au Biochemistry 2017 Only protein-protein interactions imex curation imex curation todorova-2017-2 in vitro In vitro tem transmission electron microscopy tem predetermined predetermined participant predetermined Todorova N., Zlatic CO., Mao Y., Yarovsky I., Howlett GJ., Gooley PR., Griffin MD. Samples stained with potassium phosphotungstate. Human apolipoprotein C-II was found to have a Uniprot identifier of PRO_0000002024 by mutation mapping. mgriffin@unimelb.edu.au Biochemistry 2017 Only protein-protein interactions imex curation imex curation todorova-2017-4 in vitro In vitro aggregation aggregation assay aggregation ms participant Identification by mass spectrometry ms participant Todorova N., Zlatic CO., Mao Y., Yarovsky I., Howlett GJ., Gooley PR., Griffin MD. Pellet fractions of the hybrid fibrils were further analyzed by mass spectrometry to detect the presence of each apoC-II variant by their corresponding mass. Human apolipoprotein C-II was found to have a Uniprot identifier of PRO_0000002024 by mutation mapping. mgriffin@unimelb.edu.au Biochemistry 2017 Only protein-protein interactions imex curation imex curation p02655-pro_0000002024 Proapolipoprotein C-II Apolipoprotein C2 APOC2 APC2 protein protein human Homo sapiens Human TQQPQQDEMPSPTFLTQVKESLSSYWESAKTAAQNLYEKTYLPAVDEKLRDLYSKSTAAMSTYTGIFTDQVLSVLKGEE 23 101 apoc2-apoc2-13 3649311 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3649320 3649318 direct interaction direct interaction false false false 5A Each sample showed a time-dependent increase in ThT fluorescence indicating incorporation of monomer into the preformed fibrils. Samples of preformed fibrils with added identical variant had a greater rate of change in ThT fluorescence compared to samples of preformed fibril with added K30D apoC-II variant. apoc2-apoc2-15 3649311 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P02655-PRO_0000002024:p.Asp69Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P02655-PRO_0000002024:p.Asp69Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P02655-PRO_0000002024:p.Asp69Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3649328 3649325 direct interaction direct interaction false false false 5B Each sample showed a time-dependent increase in ThT fluorescence indicating incorporation of monomer into the preformed fibrils. Samples of preformed fibrils with added identical variant had a greater rate of change in ThT fluorescence compared to samples of preformed fibril with added K30D apoC-II variant. apoc2-apoc2-16 3649311 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified whole molecule sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3649338 3649335 direct interaction direct interaction false false false 5C Each sample showed a time-dependent increase in ThT fluorescence indicating incorporation of monomer into the preformed fibrils. Samples of preformed fibrils with added identical variant had a greater rate of change in ThT fluorescence compared to samples of preformed fibril with added K30D apoC-II variant. apoc2-apoc2-17 3649311 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1A Equimolar mixtures of K30D apoC-II with WT, D69K or KDDK all showed a time- dependent increase in ThT fluorescence suggesting successful fibril formation. The maximum change in ThT fluorescence of these mixtures was higher than that of fibrils formed by WT, D69K or KDDK alone. apoc2-apoc2-18 3649311 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Asp69Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Asp69Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1B Equimolar mixtures of K30D apoC-II with WT, D69K or KDDK all showed a time- dependent increase in ThT fluorescence suggesting successful fibril formation. The maximum change in ThT fluorescence of these mixtures was higher than that of fibrils formed by WT, D69K or KDDK alone. apoc2-apoc2-19 3649311 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 1C Equimolar mixtures of K30D apoC-II with WT, D69K or KDDK all showed a time- dependent increase in ThT fluorescence suggesting successful fibril formation. The maximum change in ThT fluorescence of these mixtures was higher than that of fibrils formed by WT, D69K or KDDK alone. apoc2-apoc2-7 3649312 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Asp69Lys 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 3-3 D69K+K30D apoc2-apoc2-8 3649312 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 3-2 WT+K30D apoc2-apoc2-9 3649312 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false P02655-PRO_0000002024:p.Asp69Lys 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 P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] 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 ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false P02655-PRO_0000002024:p.Asp69Lys 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 P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 3-1 WT, D69K, and KDDK apoC-II show little protein in the supernatant fraction, and a distinct band in the pellet fraction, indicating that the majority of protein was incorporated into fibrils. ApoC-II variant K30D shows a strong band in the supernatant fraction and little protein in the pellet fraction, confirming that K30D apoC-II does not form fibrils. apoc2-apoc2-10 3649312 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp 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 3-4 KDDK+K30D apoc2-apoc2-4 3649313 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp 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 2A WT+K30D apoc2-apoc2-5 3649313 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp 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 2B KDDK+K30D apoc2-apoc2-6 3649313 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Asp69Lys mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp 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 2C D69K+K30D apoc2-apoc2-11 3649314 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 4A WT+K30D apoc2-apoc2-12 3649314 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Asp69Lys 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 4B D69K+K30D apoc2-apoc2-14 3649314 N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.[Lys30Asp;Asp69Lys] mutation mutation certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3649315 unspecified role unspecified role neutral component neutral component purified purified P02655-PRO_0000002024:p.Lys30Asp 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 4C KDDK+K30D