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 Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. FEBS J. (1742-464X) 2012 Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. in vitro In vitro dls dynamic light scattering dls predetermined predetermined participant predetermined Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. FEBS J. (1742-464X) 2012 Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. FEBS J. (1742-464X) 2012 Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel predetermined predetermined participant predetermined Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. FEBS J. (1742-464X) 2012 Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. FEBS J. (1742-464X) 2012 Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. FEBS J. (1742-464X) 2012 Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. FEBS J. (1742-464X) 2012 Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. in vitro In vitro saxs x ray scattering saxs predetermined predetermined participant predetermined Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. FEBS J. (1742-464X) 2012 Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. Functional and structural studies of the disulfide isomerase DsbC from the plant pathogen Xylella fastidiosa reveals a redox-dependent oligomeric modulation in vitro. FEBS J. (1742-464X) 2012 Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. in vitro In vitro crosslink cross-linking study crosslink western blot western blot Immuno blot Immuno blot Santos CA., Toledo MA., Trivella DB., Beloti LL., Schneider DR., Saraiva AM., Crucello A., Azzoni AR., Souza AA., Aparicio R., Souza AP. FEBS J. (1742-464X) 2012 q9pdf5_xylfa Thiol:disulfide interchange protein XF_1424 protein protein xylfa-1 Xylella fastidiosa (strain 9a5c) MYRLIVALLLGIVTLTACAQPSQPSSNKTSTTPTNATPANVQQRVTAALNTLDPDFKPDYIGAAPFPGFREVVVSGQILYVSDNGRYLIQAQPYDIQNKKFAVSEGLLSYRRTQLATVPQSQRIVFAPKNPQYTISVFTDIECGYCRKLHSEIAELNKQGIAVEYLAFPRMGLGSQDYKDMVSVWCAADRKQVLTEAKAGKQIQNKNCNNPVALEYSLGQRIGVNGTPAIFAPDGTQLGGYLPPEKLRALLDKLAAATAKGG mint C653EC9ABA07A259 q9pdf5 3 XfDsbC 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction f3b mint q9pdf5-1 1 XfDsbC 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction t1 mint We also used DLS analysis to assess the redox-dependent oligomeric modulation of XfDsbC. The results of this analysis showed that both the non-reduced and reduced condition, XfDsbC showed a low index of polydispersity (%Pd; 9.5 and 5.8, respectively) (Table 1). In addition, the molecule in solution in the non-reduced condition presents a hydrodynamic diameter (Dh) of 4.6 nm and differs from observed in the reduced condition which has a Dh of 3.8 nm, an indication of the presence of a high oligomer form of XfDsbC in the non-reduced condition. q9pdf5-1 1 XfDsbC 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 4.0 direct interaction direct interaction t1 mint We also used DLS analysis to assess the redox-dependent oligomeric modulation of XfDsbC. The results of this analysis showed that both the non-reduced and reduced condition, XfDsbC showed a low index of polydispersity (%Pd; 9.5 and 5.8, respectively) (Table 1). In addition, the molecule in solution in the non-reduced condition presents a hydrodynamic diameter (Dh) of 4.6 nm and differs from observed in the reduced condition which has a Dh of 3.8 nm, an indication of the presence of a high oligomer form of XfDsbC in the non-reduced condition. q9pdf5-1 4 XfDsbC 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 4.0 direct interaction direct interaction f4 mint The XfDsbCN-Red molecular mass obtained in the absence of TCEP was estimated to be 117 kDa, suggesting that, in the absence of reducing agent, an XfDsbC tetramer is formed in solution. q9pdf5-1 4 XfDsbC 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction f4 mint The Guinier analysis revealed a radius of gyration (Rg) of 31.2 Å. XfDsbCRed Kratky plots were also constructed (Fig. 4b) and display a well-defined maximum, which is expected for the native state of a compact globular protein. The molecular mass of the scattering particles was estimated to be 66 kDa by the SAXS MoW software [25]. This suggests that XfDsbCRed is a dimer in solution, similar to other DsbC proteins [18,26]. q9pdf5-1 5 XfDsbC 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 4.0 direct interaction direct interaction f3d mint higher-order oligomeric species that appear to correspond to tetramers were observed in vitro in the non-reduced condition of XfDsbC (Fig. 3d, lanes 1-4) after chemical crosslinking with glutaraldehyde, along with dimeric and monomeric species (Fig. 3d, lanes 2-4). However, only dimeric forms, in addition to monomeric species were observed in the reduced condition (Fig. 3d, lanes 5-8). These slow-migrating species were not observed in the absence of glutaraldehyde (Fig. 3d, lane 1 and 5). q9pdf5-1 5 XfDsbC 6 western blot western blot Immuno blot Immuno blot predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction f3d mint higher-order oligomeric species that appear to correspond to tetramers were observed in vitro in the non-reduced condition of XfDsbC (Fig. 3d, lanes 1-4) after chemical crosslinking with glutaraldehyde, along with dimeric and monomeric species (Fig. 3d, lanes 2-4). However, only dimeric forms, in addition to monomeric species were observed in the reduced condition (Fig. 3d, lanes 5-8). These slow-migrating species were not observed in the absence of glutaraldehyde (Fig. 3d, lane 1 and 5). q9pdf5-1 2 XfDsbC 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction f3a mint Under non-denaturing conditions, XfDsbC samples exhibited the same gel migration profile in the presence and absence of tris(2-carboxyethyl)phosphine (TCEP). This profile was more similar to that of conalbumin (75 kDa) than that of aldolase (158 kDa) (Fig. 3a).