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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Glutaredoxin-like protein (GLP)-a novel bacteria sulfurtransferase that protects cells against cyanide and oxidative stresses

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Author(s):
de Paula, Carla Peres [1] ; dos Santos, Melina Cardoso [1] ; Tairum, Carlos A. [1] ; Breyer, Carlos Alexandre [1] ; Toledo-Silva, Guilherme [2] ; Toyama, Marcos Hikari [3] ; Mori, Gustavo Maruyama [4] ; de Oliveira, Marcos Antonio [1]
Total Authors: 8
Affiliation:
[1] UNESP Sao Paulo State Univ, Biosci Inst, Lab Struct Mol Biol, Sao Vicente, SP - Brazil
[2] Univ Fed Santa Catarina, Dept Biochem, Lab Biomarkers Aquat Contaminat & Immunochem, Florianopolis, SC - Brazil
[3] UNESP Sao Paulo State Univ, Biosci Inst, Lab Funct & Struct Characterizat Toxins Venomous, Sao Vicente, SP - Brazil
[4] UNESP Sao Paulo State Univ, Biosci Inst, Mol Ecol Lab, Sao Vicente, SP - Brazil
Total Affiliations: 4
Document type: Journal article
Source: Applied Microbiology and Biotechnology; v. 104, n. 12 APR 2020.
Web of Science Citations: 0
Abstract

The pathogen Xylella fastidiosa belongs to the Xanthomonadaceae family, a large group of Gram-negative bacteria that cause diseases in many economically important crops. A predicted gene, annotated as glutaredoxin-like protein (glp), was found to be highly conserved among the genomes of different genera within this family and highly expressed in X. fastidiosa. Analysis of the GLP protein sequences revealed three protein domains: one similar to monothiol glutaredoxins (Grx), an Fe-S cluster and a thiosulfate sulfurtransferase/rhodanese domain (Tst/Rho), which is generally involved in sulfur metabolism and cyanide detoxification. To characterize the biochemical properties of GLP, we expressed and purified the X. fastidiosa recombinant GLP enzyme. Grx activity and Fe-S cluster formation were not observed, while an evaluation of Tst/Rho enzymatic activity revealed that GLP can detoxify cyanide and transfer inorganic sulfur to acceptor molecules in vitro. The biological activity of GLP relies on the cysteine residues in the Grx and Tst/Rho domains (Cys(33) and Cys(266), respectively), and structural analysis showed that GLP and GLP(C266S) were able to form high molecular weight oligomers (> 600 kDa), while replacement of Cys(33) with Ser destabilized the quaternary structure. In vivo heterologous enzyme expression experiments in Escherichia coli revealed that GLP can protect bacteria against high concentrations of cyanide and hydrogen peroxide. Finally, phylogenetic analysis showed that homologous glp genes are distributed across Gram-negative bacterial families with conservation of the N- to C-domain order. However, no eukaryotic organism contains this enzyme. Altogether, these results suggest that GLP is an important enzyme with cyanide-decomposing and sulfurtransferase functions in bacteria, whose presence in eukaryotes we could not observe, representing a promising biological target for new pharmaceuticals. (AU)

FAPESP's process: 13/16192-6 - Role analysis of Thr44 of Tsa1 from Saccharomyces cerevisiae in the hydroperoxide catalysis, interaction with Trx, oligomerization and overoxidation processes.
Grantee:Melina Cardoso dos Santos
Support type: Scholarships in Brazil - Scientific Initiation
FAPESP's process: 13/07937-8 - Redoxome - Redox Processes in Biomedicine
Grantee:Ohara Augusto
Support type: Research Grants - Research, Innovation and Dissemination Centers - RIDC
FAPESP's process: 17/19942-7 - Search for inhibitors of the peroxirredoxin system from pathogens and humans
Grantee:Marcos Antonio de Oliveira
Support type: Regular Research Grants
FAPESP's process: 17/06263-4 - Evaluation of bioactive molecules as inhibitors for atioxidant system of typical 2-Cys peroxiredoxins (AhpC) in baceria
Grantee:Melina Cardoso dos Santos
Support type: Scholarships in Brazil - Doctorate
FAPESP's process: 11/13500-6 - Investigation of the molecular determinants involved in the interaction with substrates of Tsa1 and Tsa2 of Saccharomyces cerevisiae
Grantee:Carlos Alexandre Breyer
Support type: Scholarships in Brazil - Doctorate
FAPESP's process: 17/20291-0 - Characterization of the proinflammatory activity of a new serine protease (cdtsp-2) purified from the total venom of Crotalus durissus terrificus
Grantee:Marcos Hikari Toyama
Support type: Regular Research Grants
FAPESP's process: 10/00172-8 - Investigation of structural transitions and reactivity towards TSA1 (thiol specific antioxidant protein 1) peroxides of Saccharomyces cerevisiae
Grantee:Carlos Abrunhosa Tairum Junior
Support type: Scholarships in Brazil - Doctorate (Direct)
FAPESP's process: 10/16827-3 - functional and Structural characterization of Glurho protein from Xylella fastidiosa.
Grantee:Carla Peres de Paula
Support type: Scholarships in Brazil - Scientific Initiation