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Kinetic characterization and search for inhibitors of Ohr (Organic Hydroperoxide Resistance Protein) from Xylella fastidiosa

Grant number: 08/07971-3
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): March 01, 2009
Effective date (End): February 28, 2011
Field of knowledge:Biological Sciences - Biochemistry - Biochemistry of Microorganisms
Principal Investigator:Luis Eduardo Soares Netto
Grantee:Thiago Geronimo Pires Alegria
Host Institution: Instituto de Biociências (IB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Associated research grant:07/58147-6 - Biological aspects of thiols: protein structure, antioxidant defense, cell signaling and redox states, AP.TEM


Reactive Oxygen Species can be generated by plants as a response against a pathogen invasion intending to eliminate it. On the other hand, during the evolution, pathogens had developed some defenses against oxidative stress. In all these kind of defenses we can highlight the protein Ohr (Organic Hydroperoxide Resistance), a peroxidase, thiol dependant, that have a reactive cysteine acting preferentially against organic peroxides and until now, it was only found in prokaryotes. Due to its presence only in bacteria and to show unique biochemical and structural functions, Ohr is a potential target for the creation of bactericide drugs. Xylella fastidiosa is a good example of a target organism for this kind of treatment. This bacterium can infect the xylem of citric plants in general and it is responsible for the CVC (Citrus Variegated Chlorosis), a disease that affects the whole plant and causes large losses in production. After the analysis of this context, this project has the objective to perform the kinetic characterization of the enzyme Ohr from Xylella fastidosa, searching for preferential substrates and inhibitors. According to the kinetic characterization, we are analyzing the enzymatic activity of Ohr using different peroxides derived from long chain fatty acids synthesized by the group of Prof. Dr. Paolo Di Mascio and Prof. Dra. Sayuri Miyamoto (IQ-USP). The search for possible inhibitors will be performed using models of virtual screening that use a known drug database and extracts from BIOprospecTA. In a second level, we want to achieve bacterial survival tests under oxidative stress in the presence of these possible inhibitors of Ohr activity. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
ALEGRIA, THIAGO G. P.; MEIRELES, DIOGO A.; CUSSIOL, JOSE R. R.; HUGO, MARTIN; TRUJILLO, MADIA; DE OLIVEIRA, MARCOS ANTONIO; MIYAMOTO, SAYURI; QUEIROZ, RAPHAEL F.; VALADARES, NAPOLEAO FONSECA; GARRATT, RICHARD C.; et al. Ohr plays a central role in bacterial responses against fatty acid hydroperoxides and peroxynitrite. Proceedings of the National Academy of Sciences of the United States of America, v. 114, n. 2, p. E132-E141, . (13/07937-8, 12/21722-1, 08/07971-3)
GOMES, FERNANDO; PALMA, FLAVIO ROMERO; BARROS, MARIO H.; TSUCHIDA, EDUARDO T.; TURANO, HELENA G.; ALEGRIA, THIAGO G. P.; DEMASI, MARILENE; NETTO, LUIS E. S.. Proteolytic cleavage by the inner membrane peptidase (IMP) complex or Oct1 peptidase controls the localization of the yeast peroxiredoxin Prx1 to distinct mitochondrial compartments. Journal of Biological Chemistry, v. 292, n. 41, p. 17011-17024, . (12/21722-1, 13/07937-8, 08/07971-3)
PICCIRILLO, ERIKA; ALEGRIA, THIAGO G. P.; DISCOLA, KAREN F.; CUSSIOL, JOSE A. R. R.; DOMINGOS, RENATO M.; DE OLIVEIRA, MARCOS A.; DE REZENDE, LEANDRO; NETTO, LUIS E. S.; AMARAL, ANTONIA T-DO. Structural insights on the efficient catalysis of hydroperoxide reduction by Ohr: Crystallographic and molecular dynamics approaches. PLoS One, v. 13, n. 5, . (14/01614-5, 12/06633-2, 08/07971-3, 05/50056-6, 16/12392-9, 13/07937-8)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
ALEGRIA, Thiago Geronimo Pires. Kinetic characterization and search for inhibitors of Ohr (Organic Hydroperoxide Resistance protein) from Xylella fastidiosa. 2012. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Biociências (IBIOC/SB) São Paulo.

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