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Functional and structural studies of the disulfide isomerase D sb C from the plant pathogen X ylella fastidiosa reveals a redox‐dependent oligomeric modulation in vitro
Author(s) -
Santos Clelton A.,
Toledo Marcelo A. S.,
Trivella Daniela B. B.,
Beloti Lilian L.,
Schneider Dilaine R. S.,
Saraiva Antonio M.,
Crucello Aline,
Azzoni Adriano R.,
Souza Alessandra A.,
Aparicio Ricardo,
Souza Anete P.
Publication year - 2012
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2012.08743.x
Subject(s) - xylella fastidiosa , periplasmic space , biofilm , chemistry , biochemistry , protein disulfide isomerase , isomerase , biophysics , bacteria , redox , oxidative folding , escherichia coli , enzyme , biology , organic chemistry , genetics , gene
X ylella fastidiosa is a G ram‐negative bacterium that grows as a biofilm inside the xylem vessels of susceptible plants and causes several economically relevant crop diseases. In the present study, we report the functional and low‐resolution structural characterization of the X . fastidiosa disulfide isomerase D sb C ( X f D sb C ). D sb C is part of the disulfide bond reduction/isomerization pathway in the bacterial periplasm and plays an important role in oxidative protein folding. In the present study, we demonstrate the presence of X f D sb C during different stages of X . fastidiosa biofilm development. X f D sb C was not detected during X . fastidiosa planktonic growth; however, after administering a sublethal copper shock, we observed an overexpression of X f D sb C that also occurred during planktonic growth. These results suggest that X . fastidiosa can use X f D sb C in vivo under oxidative stress conditions similar to those induced by copper. In addition, using dynamic light scattering and small‐angle X ‐ray scattering, we observed that the oligomeric state of X f D sb C in vitro may be dependent on the redox environment. Under reducing conditions, X f D sb C is present as a dimer, whereas a putative tetrameric form was observed under nonreducing conditions. Taken together, our findings demonstrate the overexpression of X f D sb C during biofilm formation and provide the first structural model of a bacterial disulfide isomerase in solution. Structured digital abstractXfDsbC and XfDsbC bind by x ray scattering (View Interaction: 1 , 2 ) XfDsbC and XfDsbC bind by molecular sieving ( View interaction ) XfDsbC and XfDsbC bind by comigration in non denaturing gel electrophoresis ( View interaction ) XfDsbC and XfDsbC bind by cross‐linking study (View Interaction: 1 , 2 ) XfDsbC and XfDsbC bind by dynamic light scattering (View Interaction: 1 , 2 )