MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it http://mint.bio.uniroma2.it/mint xiumin-2013-1 Isobavachalcone and bavachinin from Psoraleae Fructus modulate Aβ42 aggregation process through different mechanisms in vitro. in vitro In vitro comigration in sds comigration in sds page comigration in sds weight by comassie molecular weight estimation by coomasie staining weight by comassie Chen X., Yang Y., Zhang Y. Two major bands corresponding to monomers and low molecular weight conformers (trimers/tetramers) were observed in samples before incubation (Fig. 3A). After 6 h incubation, high molecular weight aggregates markedly accumulated in the samples. Addition of IBC greatly prevented the formation of high molecular weight aggregates, further confirmed its anti-aggregation effect (Fig. 3A, 3B). zytao@bjmu.edu.cn FEBS Lett. (0014-5793) 2013 Only protein-protein interactions imex curation xiumin-2013-2 Isobavachalcone and bavachinin from Psoraleae Fructus modulate Aβ42 aggregation process through different mechanisms in vitro. in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Chen X., Yang Y., Zhang Y. Two major bands corresponding to monomers and low molecular weight conformers (trimers/tetramers) were observed in samples before incubation (Fig. 3A). After 6 h incubation, high molecular weight aggregates markedly accumulated in the samples. Addition of IBC greatly prevented the formation of high molecular weight aggregates, further confirmed its anti-aggregation effect (Fig. 3A, 3B). zytao@bjmu.edu.cn FEBS Lett. (0014-5793) 2013 Only protein-protein interactions imex curation xiumin-2013-3 Isobavachalcone and bavachinin from Psoraleae Fructus modulate Aβ42 aggregation process through different mechanisms in vitro. in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay western blot western blot Immuno blot Immuno blot Chen X., Yang Y., Zhang Y. Two major bands corresponding to monomers and low molecular weight conformers (trimers/tetramers) were observed in samples before incubation (Fig. 3A). After 6 h incubation, high molecular weight aggregates markedly accumulated in the samples. Addition of IBC greatly prevented the formation of high molecular weight aggregates, further confirmed its anti-aggregation effect (Fig. 3A, 3B). zytao@bjmu.edu.cn FEBS Lett. (0014-5793) 2013 Only protein-protein interactions imex curation xiumin-2013-4 Isobavachalcone and bavachinin from Psoraleae Fructus modulate Aβ42 aggregation process through different mechanisms in vitro. in vitro In vitro atomic force microsc atomic force microscopy AFM AFM atomic force microsc predetermined predetermined participant predetermined Chen X., Yang Y., Zhang Y. Two major bands corresponding to monomers and low molecular weight conformers (trimers/tetramers) were observed in samples before incubation (Fig. 3A). After 6 h incubation, high molecular weight aggregates markedly accumulated in the samples. Addition of IBC greatly prevented the formation of high molecular weight aggregates, further confirmed its anti-aggregation effect (Fig. 3A, 3B). zytao@bjmu.edu.cn FEBS Lett. (0014-5793) 2013 Only protein-protein interactions imex curation xiumin-2013-5 Isobavachalcone and bavachinin from Psoraleae Fructus modulate Aβ42 aggregation process through different mechanisms in vitro. yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Chen X., Yang Y., Zhang Y. Two major bands corresponding to monomers and low molecular weight conformers (trimers/tetramers) were observed in samples before incubation (Fig. 3A). After 6 h incubation, high molecular weight aggregates markedly accumulated in the samples. Addition of IBC greatly prevented the formation of high molecular weight aggregates, further confirmed its anti-aggregation effect (Fig. 3A, 3B). zytao@bjmu.edu.cn FEBS Lett. (0014-5793) 2013 Only protein-protein interactions imex curation a4_human Amyloid-beta precursor protein APP A4 AD1 Alzheimer disease amyloid protein ABPP PreA4 Cerebral vascular amyloid peptide Protease nexin-II APPI Amyloid precursor protein Amyloid-beta A4 protein Alzheimer disease amyloid A4 protein homolog Amyloid-beta (A4) precursor protein protein protein human Homo sapiens Human MLPGLALLLLAAWTARALEVPTDGNAGLLAEPQIAMFCGRLNMHMNVQNGKWDSDPSGTKTCIDTKEGILQYCQEVYPELQITNVVEANQPVTIQNWCKRGRKQCKTHPHFVIPYRCLVGEFVSDALLVPDKCKFLHQERMDVCETHLHWHTVAKETCSEKSTNLHDYGMLLPCGIDKFRGVEFVCCPLAEESDNVDSADAEEDDSDVWWGGADTDYADGSEDKVVEVAEEEEVAEVEEEEADDDEDDEDGDEVEEEAEEPYEEATERTTSIATTTTTTTESVEEVVREVCSEQAETGPCRAMISRWYFDVTEGKCAPFFYGGCGGNRNNFDTEEYCMAVCGSAMSQSLLKTTQEPLARDPVKLPTTAASTPDAVDKYLETPGDENEHAHFQKAKERLEAKHRERMSQVMREWEEAERQAKNLPKADKKAVIQHFQEKVESLEQEAANERQQLVETHMARVEAMLNDRRRLALENYITALQAVPPRPRHVFNMLKKYVRAEQKDRQHTLKHFEHVRMVDPKKAAQIRSQVMTHLRVIYERMNQSLSLLYNVPAVAEEIQDEVDELLQKEQNYSDDVLANMISEPRISYGNDALMPSLTETKTTVELLPVNGEFSLDDLQPWHSFGADSVPANTENEVEPVDARPAADRGLTTRPGSGLTNIKTEEISEVKMDAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIATVIVITLVMLKKKQYTSIHHGVVEVDAAVTPEERHLSKMQQNGYENPTYKFFEQMQN app-1 1427389 n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 4.0 direct interaction direct interaction false false false f3 app-1 1427389 n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro Stoichiometry: 3.0 direct interaction direct interaction false false false f3 Two major bands corresponding to monomers and low molecular weight conformers (trimers/tetramers) were observed in samples before incubation (Fig. 3A). After 6 h incubation, high molecular weight aggregates markedly accumulated in the samples. Addition of IBC greatly prevented the formation of high molecular weight aggregates, further confirmed its anti-aggregation effect (Fig. 3A, 3B). app-app-5 1427389 n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro direct interaction direct interaction false false false f3 Two major bands corresponding to monomers and low molecular weight conformers (trimers/tetramers) were observed in samples before incubation (Fig. 3A). After 6 h incubation, high molecular weight aggregates markedly accumulated in the samples. Addition of IBC greatly prevented the formation of high molecular weight aggregates, further confirmed its anti-aggregation effect (Fig. 3A, 3B). app-app-6 1427390 n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro direct interaction direct interaction false false false f2b f5a f5b To further confirm whether PFE and the compounds can inhibit aggregation, a standard ThT fluorescence assay was performed. ThT is known to specifically bind to fibrillar conformers and generate a unique fluorescence spectrum peaking near 490 nm. As shown in Fig. 2B, both the positive control EGCG and PFE significantly reduced ThT fluorescence, indicating they inhibited formation of fibrillar aggregates.After 7 days of incubation, alone formed significant amount of aggregates that can be pelleted by centrifugation and recognized by ThT indicated by the increase of fluorescence at 490 nm (Fig. 5A). app-app-7 1427391 n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro direct interaction direct interaction false false false f4 To elucidate which pathway of aggregation was modulated by IBC and BCN, we performed dot blot assay using oligomer-specific A11 antibody and fibril-specific OC antibody. OC antibody recognizes fibrillar intermediates and mature amyloid fibrils that are less toxic, ThT-positive and beta-sheet-rich structures [21,23]. When was incubated alone, OC positive conformers accumulated significantly as time progressed (Fig 4). However, this accumulation was almost completely stopped when either BCN or IBC was added at a molar ratio of 5:1 (BCN or IBC to ) and the inhibition by either compound was dose-dependent (Fig 4). This demonstrated both compounds inhibited fibrillization process app-app-8 1427392 n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro n/a Abeta42 1427394 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false in vitro In vitro direct interaction direct interaction false false false f6a f6b However, when examined by atomic force microscopy, fibrils formed from wt moPrP and the S170N/N174T variant appeared morphologically very similar. In both cases, fibrils were of variable length, and AFM images indicated the presence of both individual protofilaments as well as thicker (higher) structures resulting from lateral association of these protofilaments (Fig. 3 A and B). app-app-10 1427393 n/a Abeta42 1427394 unspecified role unspecified role prey prey living cell living cell tag fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false n/a Abeta42 1427394 unspecified role unspecified role bait bait living cell living cell binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tag fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false f2a the yeast growth in high-stringency SD4- media is completely dependent on self interaction thus was used as a read out for evaluation of inhibitors. Toxicity dependent false-positive effects were identified by a parallel test replacing SD4- with complete SD media. As shown in Fig. 2A, PFE (10 ug/ml) and curcumin (150 uM), a known inhibitor, significantly reduced the yeast growth in SD4- medium, indicating that they strongly inhibit dimerization. Among the eight test compounds, IBC (3 uM) and BCN (30 uM) were found to be active at non-toxic concentrations.