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 Orientation of the amino-terminal domain of ClpB affects the disaggregation of the protein. Orientation of the amino-terminal domain of ClpB affects the disaggregation of the protein. FEBS J. (1742-464X) 2012 imex curation Mizuno S., Nakazaki Y., Yoshida M., Watanabe YH. Only protein-protein interactions in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Mizuno S., Nakazaki Y., Yoshida M., Watanabe YH. FEBS J. (1742-464X) 2012 Only protein-protein interactions imex curation Orientation of the amino-terminal domain of ClpB affects the disaggregation of the protein. Orientation of the amino-terminal domain of ClpB affects the disaggregation of the protein. FEBS J. (1742-464X) 2012 imex curation Mizuno S., Nakazaki Y., Yoshida M., Watanabe YH. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Mizuno S., Nakazaki Y., Yoshida M., Watanabe YH. FEBS J. (1742-464X) 2012 Only protein-protein interactions imex curation cask_bovin Kappa-casein CSN3 CSN10 CSNK protein protein bovin Bos taurus (Bovine) Bovine MMKSFFLVVTILALTLPFLGAQEQNQEQPIRCEKDERFFSDKIAKYIPIQYVLSRYPSYGLNYYQQKPVALINNQFLPYPYYAKPAAVRSPAQILQWQVLSNTVPAKSCQAQPTTMARHPHPHLSFMAIPPKKNQDKTEIPTINTIASGEPTSTPTTEAVESTVATLEDSPEVIESPPEINTVQVTSTAV mint F12D8310C3B93EDA clpb_thet8 Chaperone protein ClpB clpB TTHA1487 protein protein thet8 Thermus thermophilus (strain HB8 / ATCC 27634 / DSM 579) MNLERWTQAAREALAQAQVLAQRMKHQAIDLPHLWAVLLKDERSLAWRLLEKAGADPKALKELQERELARLPKVEGAEVGQYLTSRLSGALNRAEALMEELKDRYVAVDTLVLALAEATPGLPGLEALKGALKELRGGRTVQTEHAESTYNALEQYGIDLTRLAAEGKLDPVIGRDEEIRRVIQILLRRTKNNPVLIGEPGVGKTAIVEGLAQRIVKGDVPEGLKGKRIVSLQMGSLLAGAKYRGEFEERLKAVIQEVVQSQGEVILFIDELHTVVGAGKAEGAVDAGNMLKPALARGELRLIGATTLDEYREIEKDPALERRFQPVYVDEPTVEETISILRGLKEKYEVHHGVRISDSAIIAAATLSHRYITERRLPDKAIDLIDEAAARLRMALESAPEEIDALERKKLQLEIEREALKKEKDPDSQERLKAIEAEIAKLTEEIAKLRAEWEREREILRKLREAQHRLDEVRREIELAERQYDLNRAAELRYGELPKLEAEVEALSEKLRGARFVRLEVTEEDIAEIVSRWTGIPVSKLLEGEREKLLRLEEELHKRVVGQDEAIRAVADAIRRARAGLKDPNRPIGSFLFLGPTGVGKTELAKTLAATLFDTEEAMIRIDMTEYMEKHAVSRLIGAPPGYVGYEEGGQLTEAVRRRPYSVILFDEIEKAHPDVFNILLQILDDGRLTDSHGRTVDFRNTVIILTSNLGSPLILEGLQKGWPYERIRDEVFKVLQQHFRPEFLNRLDEIVVFRPLTKEQIRQIVEIQLSYLRARLAEKRISLELTEAAKDFLAERGYDPVFGARPLRRVIQRELETPLAQKILAGEVKEGDRVQVDVGPAGLVFAVPARVEA DIP protein Q9RA63 original sequence version: 92 mint Part of a stress-induced multi-chaperone system, it is involved in the recovery of the cell from heat-induced damage, in cooperation with dnaK, dnaJ and grpE. Acts before dnaK, in the processing of protein aggregates. Protein binding stimulates the ATPase activity; ATP hydrolysis unfolds the denatured protein aggregates, which probably helps expose new hydrophobic binding sites on the surface of clpB-bound aggregates, contributing to the solubilization and refolding of denatured protein aggregates by dnaK FB9A39BB040363E0 clpb-1 2 TClpB 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 6.0 direct interaction direct interaction f3 mint In this condition, NXin, NXout, and R23C were eluted at the same position as wild-type TClpB (Fig. 3), and the oxidized forms of these mutants were confirmed to form a stable hexamer. In addition, the stability was not influenced by the pre-incubation with κ-casein and adenosine 5′-O-(thiotriphosphate) (ATPγS) (Fig. 3). clpb-csn3 1 K-casein 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.6 TClpB 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 6.1 direct interaction direct interaction f4 mint Binding of soluble denatured protein by TClpB mutants. Wild-type and mutant TClpB were pre-incubated with ATPγS at 55 °C and formed stable hexamers. Various concentrations of these hexamers were mixed with FITC-labeled κ-casein (FITC-κ-casein), and the fluorescence polarities of FITC were measured at the same temperature (Fig. 4). The apparent dissociation constants (Kd) of wild-type and mutant TClpB hexamer for FITC-κ-casein were calculated by fitting the data to a model in which a TClpB hexamer binds to an FITC-κ-casein. The Kd of TClpB N (0.28 μM) was 14 times higher than that of the wild type (0.02 μM). However, the Kd values of the oxidized cysteine mutants, NXin (0.02 μM), NXout (0.03 μM), and R23C (0.04 μM), were similar to those of the wild type, even in the absence of a reducing agent. clpb-csn3-1 1 K-casein 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.6 TClpB 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined 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 Stoichiometry: 6.1 direct interaction direct interaction f4 mint Binding of soluble denatured protein by TClpB mutants. Wild-type and mutant TClpB were pre-incubated with ATPγS at 55 °C and formed stable hexamers. Various concentrations of these hexamers were mixed with FITC-labeled κ-casein (FITC-κ-casein), and the fluorescence polarities of FITC were measured at the same temperature (Fig. 4). The apparent dissociation constants (Kd) of wild-type and mutant TClpB hexamer for FITC-κ-casein were calculated by fitting the data to a model in which a TClpB hexamer binds to an FITC-κ-casein. The Kd of TClpB N (0.28 μM) was 14 times higher than that of the wild type (0.02 μM). However, the Kd values of the oxidized cysteine mutants, NXin (0.02 μM), NXout (0.03 μM), and R23C (0.04 μM), were similar to those of the wild type, even in the absence of a reducing agent. clpb-csn3-2 1 K-casein 3 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.6 TClpB 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 6.1 direct interaction direct interaction f4 mint Binding of soluble denatured protein by TClpB mutants. Wild-type and mutant TClpB were pre-incubated with ATPγS at 55 °C and formed stable hexamers. Various concentrations of these hexamers were mixed with FITC-labeled κ-casein (FITC-κ-casein), and the fluorescence polarities of FITC were measured at the same temperature (Fig. 4). The apparent dissociation constants (Kd) of wild-type and mutant TClpB hexamer for FITC-κ-casein were calculated by fitting the data to a model in which a TClpB hexamer binds to an FITC-κ-casein. The Kd of TClpB N (0.28 μM) was 14 times higher than that of the wild type (0.02 μM). However, the Kd values of the oxidized cysteine mutants, NXin (0.02 μM), NXout (0.03 μM), and R23C (0.04 μM), were similar to those of the wild type, even in the absence of a reducing agent.