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 anonymous-2014c-1 Structure-Function of Cyanobacterial Outer-Membrane Protein, Slr1270: Homolog of E. coli Drug Export/ Colicin Import Protein, TolC in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined waclab@purdue.edu 2014 imex curation Agarwal R., Zakharov S., Hasan S.S., Ryan C.M., Whitelegge J.P., Cramer W.A. FEBS Lett. (0014-5793) Only protein-protein interactions p74182_syny3 slr1270 protein protein syny3 Synechocystis sp. (strain PCC 6803 / Kazusa) MKSIHPLKFWSSSTLLLLLSTSVGVFLPGFSGGQGAIAVAQSVSPPDNAPSAETGNNADDSGFPLPQLPDTAPNERLNPSGNPLMFPTKPDEVDTTVRQAITLDEAIDLALRNNEQLQQAKLSLEQQEAGLMAARAALFPSLDTDFTFSRDSSAAAEATNALIANQNQTTTVNPELRSETSTNAVGNINLTYSIYAGGERSAQIAKAEQLVQNSRLQVEVVAEQTRFEATDRYYALQGADAQVAIAQASVEDASQSLRDARLLEQAGLGTRFDVLRAEGDLATANEALTRSIADQRNARRRLAQLLSVGQRVELTAADEIVEAGDWSLPLDESIVQAYKNRAELEQQLVQIEVSEQDRYIALAAIKPRVDFLANYTYQNNFDSSAGLVDGYAFAARVRWNFFDGGRAFAEARRADRQMDIAKTAFSEQRNQIRLEVEESYYTLISNKENIGSTRTNVIRFEEALRLARLRFQAGVGTQTDVINAQRDLANARGRFLQAIIGYNQSLNQLQRSISNLPNNHLFDIQP tolc_ecoli Outer membrane protein TolC tolC mtcB mukA refI Multidrug efflux pump subunit TolC Outer membrane factor TolC colE1-i toc weeA JW5503 b3035 protein protein ecoli Escherichia coli (strain K12) MKKLLPILIGLSLSGFSSLSQAENLMQVYQQARLSNPELRKSAADRDAAFEKINEARSPLLPQLGLGADYTYSNGYRDANGINSNATSASLQLTQSIFDMSKWRALTLQEKAAGIQDVTYQTDQQTLILNTATAYFNVLNAIDVLSYTQAQKEAIYRQLDQTTQRFNVGLVAITDVQNARAQYDTVLANEVTARNNLDNAVEQLRQITGNYYPELAALNVENFKTDKPQPVNALLKEAEKRNLSLLQARLSQDLAREQIRQAQDGHLPTLDLTASTGISDTSYSGSKTRGAAGTQYDDSNMGQNKVGLSFSLPIYQGGMVNSQVKQAQYNFVGASEQLESAHRSVVQTVRSSFNNINASISSINAYKQAVVSAQSSLDAMEAGYSVGTRTIVDVLDATTTLYNAKQELANARYNYLINQLNIKSALGTLNEQDLLALNNALSKPVSTNPENVAPQTPEQNAIADGYAPDSPAPVVQQTSARTTTSNGHNPFRN p74182-2 1565164 n/a Slr1270 1565165 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f3 Slr1270 fractions purified by Ni-NTA chromatography were pooled and refolded through stepwise dialysis in the presence of 0.1% UDM detergent. The refolded protein, loaded on a Superdex- 200 size exclusion FPLC column, eluted in two peaks at 8.3 mL and 10.7 mL (Fig. 3A), similar to the elution pattern of native E. coli TolC (elution curve in red), supporting correct refolding of the Slr1270. The peak at 8.3 mL corresponds to protein aggregates with Mr > 400 kDa (Fig. 3A). The peak at 10.7 mL contained trimeric Slr1270 protein (Fig. 3A) as it displayed a single band on clear-native PAGE corresponding to a trimer similar to native E. coli TolC (Fig. 3B, lanes 2 and 5). The small difference in migration is consistent with the difference in protein size (Slr1270 monomer ~ 55 kDa; E. coli TolC monomer ~ 50 kDa). However, the Slr1270 protein showed a concentration dependent aggregation (Fig. 3B, lanes 3-4). p74182-p74182-1 1565164 n/a Slr1270 1565165 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a Slr1270 1565165 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false f3 Slr1270 fractions purified by Ni-NTA chromatography were pooled and refolded through stepwise dialysis in the presence of 0.1% UDM detergent. The refolded protein, loaded on a Superdex- 200 size exclusion FPLC column, eluted in two peaks at 8.3 mL and 10.7 mL (Fig. 3A), similar to the elution pattern of native E. coli TolC (elution curve in red), supporting correct refolding of the Slr1270. The peak at 8.3 mL corresponds to protein aggregates with Mr > 400 kDa (Fig. 3A). The peak at 10.7 mL contained trimeric Slr1270 protein (Fig. 3A) as it displayed a single band on clear-native PAGE corresponding to a trimer similar to native E. coli TolC (Fig. 3B, lanes 2 and 5). The small difference in migration is consistent with the difference in protein size (Slr1270 monomer ~ 55 kDa; E. coli TolC monomer ~ 50 kDa). However, the Slr1270 protein showed a concentration dependent aggregation (Fig. 3B, lanes 3-4). tolc-1 1565164 n/a TolC 1565166 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 3.0 direct interaction direct interaction false false false f3a f3c Slr1270 fractions purified by Ni-NTA chromatography were pooled and refolded through stepwise dialysis in the presence of 0.1% UDM detergent. The refolded protein, loaded on a Superdex- 200 size exclusion FPLC column, eluted in two peaks at 8.3 mL and 10.7 mL (Fig. 3A), similar to the elution pattern of native E. coli TolC (elution curve in red), supporting correct refolding of the Slr1270. The peak at 8.3 mL corresponds to protein aggregates with Mr > 400 kDa (Fig. 3A). The peak at 10.7 mL contained trimeric Slr1270 protein (Fig. 3A) as it displayed a single band on clear-native PAGE corresponding to a trimer similar to native E. coli TolC (Fig. 3B, lanes 2 and 5). The small difference in migration is consistent with the difference in protein size (Slr1270 monomer ~ 55 kDa; E. coli TolC monomer ~ 50 kDa). However, the Slr1270 protein showed a concentration dependent aggregation (Fig. 3B, lanes 3-4).