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 Facilitated aggregation of FG nucleoporins under molecular crowding conditions. Facilitated aggregation of FG nucleoporins under molecular crowding conditions. EMBO Rep. (1469-221X) 2013 imex curation Milles S., Huy Bui K., Koehler C., Eltsov M., Beck M., Lemke EA. Only protein-protein interactions in vitro In vitro fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Milles S., Huy Bui K., Koehler C., Eltsov M., Beck M., Lemke EA. EMBO Rep. (1469-221X) 2013 Only protein-protein interactions imex curation Facilitated aggregation of FG nucleoporins under molecular crowding conditions. Facilitated aggregation of FG nucleoporins under molecular crowding conditions. EMBO Rep. (1469-221X) 2013 imex curation Milles S., Huy Bui K., Koehler C., Eltsov M., Beck M., Lemke EA. Only protein-protein interactions in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Milles S., Huy Bui K., Koehler C., Eltsov M., Beck M., Lemke EA. EMBO Rep. (1469-221X) 2013 Only protein-protein interactions imex curation Facilitated aggregation of FG nucleoporins under molecular crowding conditions. Facilitated aggregation of FG nucleoporins under molecular crowding conditions. EMBO Rep. (1469-221X) 2013 imex curation Milles S., Huy Bui K., Koehler C., Eltsov M., Beck M., Lemke EA. Only protein-protein interactions in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Milles S., Huy Bui K., Koehler C., Eltsov M., Beck M., Lemke EA. EMBO Rep. (1469-221X) 2013 Only protein-protein interactions imex curation nup49_yeast Nucleoporin NUP49/NSP49 NUP49 NSP49 YGL172W G1648 Nuclear pore protein NUP49/NSP49 protein protein yeast Saccharomyces cerevisiae Baker's yeast MFGLNKASSTPAGGLFGQASGASTGNANTGFSFGGTQTGQNTGPSTGGLFGAKPAGSTGGLGASFGQQQQQSQTNAFGGSATTGGGLFGNKPNNTANTGGGLFGANSNSNSGSLFGSNNAQTSRGLFGNNNTNNINNSSSGMNNASAGLFGSKPAGGTSLFGNTSTSSAPAQNQGMFGAKPAGTSLFGNNAGNTTTGGGLFGSKPTGATSLFGSSNNNNNNNNSNNIMSASGGLFGNQQQQLQQQPQMQCALQNLSQLPITPMTRISELPPQIRQEIEQLDQYIQKQVQISHHLKADTIDHDELIDSIPRDVAYLLKSESATSQYLKQDLKKISSFKSLIDEDLLDTQTFSVLLQQLLTPGSKISSNDLDKFFQKKIHLYEKKLEDYCRILSDIETAVNGIDTDLFGAPNNPNSTAITADLGSSEAENLLQLKTGLAAIVSTVIEEFTLFMDIAERIAVLHQKTKTLASLSI 0CA7516FF5753CA3 nu153_human Nuclear pore complex protein Nup153 Nucleoporin Nup153 153 kDa nucleoporin NUP153 protein protein human Homo sapiens Human MASGAGGVGGGGGGKIRTRRCHQGPIKPYQQGRQQHQGILSRVTESVKNIVPGWLQRYFNKNEDVCSCSTDTSEVPRWPENKEDHLVYADEESSNITDGRITPEPAVSNTEEPSTTSTASNYPDVLTRPSLHRSHLNFSMLESPALHCQPSTSSAFPIGSSGFSLVKEIKDSTSQHDDDNISTTSGFSSRASDKDITVSKNTSLPPLWSPEAERSHSLSQHTATSSKKPAFNLSAFGTLSPSLGNSSILKTSQLGDSPFYPGKTTYGGAAAAVRQSKLRNTPYQAPVRRQMKAKQLSAQSYGVTSSTARRILQSLEKMSSPLADAKRIPSIVSSPLNSPLDRSGIDITDFQAKREKVDSQYPPVQRLMTPKPVSIATNRSVYFKPSLTPSGEFRKTNQRIDNKCSTGYEKNMTPGQNREQRESGFSYPNFSLPAANGLSSGVGGGGGKMRRERTRFVASKPLEEEEMEVPVLPKISLPITSSSLPTFNFSSPEITTSSPSPINSSQALTNKVQMTSPSSTGSPMFKFSSPIVKSTEANVLPPSSIGFTFSVPVAKTAELSGSSSTLEPIISSSAHHVTTVNSTNCKKTPPEDCEGPFRPAEILKEGSVLDILKSPGFASPKIDSVAAQPTATSPVVYTRPAISSFSSSGIGFGESLKAGSSWQCDTCLLQNKVTDNKCIACQAAKLSPRDTAKQTGIETPNKSGKTTLSASGTGFGDKFKPVIGTWDCDTCLVQNKPEAIKCVACETPKPGTCVKRALTLTVVSESAETMTASSSSCTVTTGTLGFGDKFKRPIGSWECSVCCVSNNAEDNKCVSCMSEKPGSSVPASSSSTVPVSLPSGGSLGLEKFKKPEGSWDCELCLVQNKADSTKCLACESAKPGTKSGFKGFDTSSSSSNSAASSSFKFGVSSSSSGPSQTLTSTGNFKFGDQGGFKIGVSSDSGSINPMSEGFKFSKPIGDFKFGVSSESKPEEVKKDSKNDNFKFGLSSGLSNPVSLTPFQFGVSNLGQEEKKEELPKSSSAGFSFGTGVINSTPAPANTIVTSENKSSFNLGTIETKSASVAPFTCKTSEAKKEEMPATKGGFSFGNVEPASLPSASVFVLGRTEEKQQEPVTSTSLVFGKKADNEEPKCQPVFSFGNSEQTKDENSSKSTFSFSMTKPSEKESEQPAKATFAFGAQTSTTADQGAAKPVFSFLNNSSSSSSTPATSAGGGIFGSSTSSSNPPVATFVFGQSSNPVSSSAFGNTAESSTSQSLLFSQDSKLATTSSTGTAVTPFVFGPGASSNNTTTSGFGFGATTTSSSAGSSFVFGTGPSAPSASPAFGANQTPTFGQSQGASQPNPPGFGSISSSTALFPTGSQPAPPTFGTVSSSSQPPVFGQQPSQSAFGSGTTPNSSSAFQFGSSTTNFNFTNNSPSGVFTFGANSSTPAASAQPSGSGGFPFNQSPAAFTVGSNGKNVFSSSGTSFSGRKIKTAVRRRK DIP protein P49790 original sequence version: 94 D07455A691F7CD1F nup153-nup153 3 hNUP153 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain hNUP153 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction homomint mint f1a f2c An increase of ThT fluorescence by orders of magnitudes as well as the prototypical time dependence of amyloid formation showing a lag phase, a burst phase and finally a saturation phase (also illustrated in supplementary Fig S1a online) is commonly used for amyloid detection [26]. hNup153FG clearly followed both requirements (Fig 1A) in the ThT assay, and the formation of amyloids with a typical amyloid twist was also confirmed using EM (Fig 1B,C) [27]. nup49-nup49 3 yNUP49 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain yNUP49 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction sf1a homomint mint As human and yeast FG Nups share only little sequence similarity, we subjected yNup49FG to the same test, and determined the same phenotype as for hNup153FG (supplementary Fig S1 online). nup153-nup153-1 2 hNUP153 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain hNUP153 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction f1b f1c homomint mint hNup153FG clearly followed both requirements (Fig 1A) in the ThT assay, and the formation of amyloids with a typical amyloid twist was also confirmed using EM (Fig 1B,C) [27]. nup49-nup49-1 2 yNUP49 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain yNUP49 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction sf1b homomint mint As human and yeast FG Nups share only little sequence similarity, we subjected yNup49FG to the same test, and determined the same phenotype as for hNup153FG (supplementary Fig S1 online). nup153-nup153-2 1 hNUP153 5 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp unspecified role unspecified role fluorescence donor fluorescence donor purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined hNUP153 5 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction homomint mint Two batches of hNup153FG were labelled with two different dyes, such that FRET can occur between a Donor (D, Alexa488) and an Acceptor (A, Alexa594) dye only if two molecules come in close proximity. D- and A-labelled hNup153FG were mixed with serine and immediately a clear FRET signal became apparent that rose with increasing serine concentrations in a sigmoidal fashion (Fig 1D,E). This aggregation occurred even at the single-molecule level (pM protein concentrations, see Fig 1F; supplementary Fig S2 online) and is thus extremely high under molecular crowding conditions. As shown in Fig 2, we incubated saturating concentrations of Importinb together with a 1:1 mixture from separate batches of Alexa488- and Alexa594-labelled hNup153FG, and then transferred this mixture to molecular crowding conditions. No FRET signal between the dyes was observed, indicating that Importinb prevents hNup153FG molecules to come into such proximity thatthe two dyes can undergo FRET. Even if Importinb was added after aggregation was induced, it could immediately stop further increase of the FRET signal compared with a sample that did not contain the transport receptor (Fig 2B). f1d f2a f2b sf2 nup153-nup153-3 1 hNUP153 5 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp unspecified role unspecified role fluorescence donor fluorescence donor purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined hNUP153 5 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction homomint mint f1d f1f Two batches of hNup153FG were labelled with two different dyes, such that FRET can occur between a Donor (D, Alexa488) and an Acceptor (A, Alexa594) dye only if two molecules come in close proximity. D- and A-labelled hNup153FG were mixed with serine and immediately a clear FRET signal became apparent that rose with increasing serine concentrations in a sigmoidal fashion (Fig 1D,E). This aggregation occurred even at the single-molecule level (pM protein concentrations, see Fig 1F; supplementary Fig S2 online) and is thus extremely high under molecular crowding conditions. nup49-nup49-2 1 yNUP49 4 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp unspecified role unspecified role fluorescence donor fluorescence donor purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yNUP49 4 tag fluorescence tag visualisation by fluorescence tag fluorescence experimental particp experimental participant identification experimental particp unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined direct interaction direct interaction sf1c homomint mint As human and yeast FG Nups share only little sequence similarity, we subjected yNup49FG to the same test, and determined the same phenotype as for hNup153FG (supplementary Fig S1 online).