Functional and structural studies of the
disulfide isomerase DsbC from the plant
pathogen Xylella fastidiosa reveals a
redox-dependent oligomeric modulation
in vitro
Clelton A. Santos, Marcelo A. S. Toledo, Daniela B. B. Trivella,
Lilian L. Beloti, Dilaine R. S. Schneider, Antonio M. Saraiva,
Aline Crucello, Adriano R. Azzoni, Alessandra A. Souza, Ricardo Aparicio
and Anete P. Souza
DOI: 10.1111/j.1742-4658.2012.08743.x
Functional and Structural Studies of the Disulfide Isomerase XfDsbC from the Plant
Pathogen Xylella fastidiosa Reveals a Redox-dependent Oligomeric Modulation in vitro
Clelton A. Santos1, Marcelo A.S. Toledo1, Daniela B.B. Trivella2, Lilian L. Beloti1, Dilaine
R.S. Schneider1, Antonio M. Saraiva1, Aline Crucello1, Adriano R. Azzoni1,3, Alessandra A.
Souza4, Ricardo Aparicio2 and Anete P. Souza1,5,*
1
Centro de Biologia Molecular e Engenharia Genética, Universidade Estadual de Campinas,
Campinas, SP, Brazil
2
Laboratório de Biologia Estrutural e Cristalografia, Instituto de Química, Universidade
Estadual de Campinas, Campinas, SP, Brazil
3
Departamento de Engenharia Química, Escola Politécnica, Universidade de São Paulo, São
Paulo, SP, Brazil
4
Centro APTA Citros Sylvio Moreira/IAC, Rodovia Anhanguera Km 158, Cordeirópolis, SP,
Brazil
5
Departamento de Biologia Vegetal, Instituto de Biologia, Universidade Estadual de
Campinas, Campinas, SP, Brazil
Supplementary Figures S1-S3
*Address correspondence to: Anete P. Souza, Centro de Biologia Molecular e Engenharia
Genética (CBMEG), Universidade Estadual de Campinas, Rua Candido Rondon 400, Cidade
Universitária “Zeferino Vaz”, CP 6010, 13083-875, Campinas, São Paulo, Brazil, Tel. and
Fax: +55 19 3521-1089; E-mail: [email protected]
Supplemental Figure S1. ClustalW2 alignment of DsbC sequences from Xylella fastidiosa
strain 9a5c (XfDsbC), Xanthomonas campestris pv. campestris (XcDsbC), Erwinia
chrysanthemi (ErcDsbC) and Escherichia coli (EcDsbC). The regions that are fully conserved
in the four sequences (black) and the residues that are strongly similar between the sequences
(gray) are highlighted. The oxi-redox switch, which is composed of the CGYC motif, is
indicated in red in all sequences.
Supplemental Figure S2. The inverted XfDsbCRed dimer. Three-dimensional atomic model
for XfDsbC dimer was generated as an output from BUNCH software. The low-resolution
data obtained limits the interpretation of the correct XfDsbCRed dimeric structure. Therefore,
a second possible XfDsbCRed dimer is also provided. The same low-resolution protein
envelope constructed for the XfDsbCRed dimer displayed in Fig. 5A is shown here as semitransparent whitish spheres superimposed onto the inverted XfDsbCRed dimer atomic model.
The middle and bottom views are rotated clockwise by 90° around the x- and y-axes,
respectively. The figure was generated with PyMOL software (Schrodinger, LLC).
A
B
Supplemental Figure S3. Possible tetrameric assemblies of (A) EcDsbC (PDB ID 1EEJ) and
(B) HiDsbC (PDB ID 1TDJ). These oligomers were observed n the crystal structures by
applying symmetric operations. The symmetric molecules and figures were generated with
PyMOL software (Schrodinger, LLC).