| Full text | |
| 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 Opportunities: | Scholarships in Brazil - Scientific Initiation |
| FAPESP's process: | 13/07937-8 - Redoxome - Redox Processes in Biomedicine |
| Grantee: | Ohara Augusto |
| Support Opportunities: | 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 Opportunities: | 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 Opportunities: | 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 Opportunities: | 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 Opportunities: | 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 Opportunities: | 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 Opportunities: | Scholarships in Brazil - Scientific Initiation |