Natália Pinto de Almeida1,2, Domingos Ferreira Mélo Neto3, Gabriel Reis Alves Carneiro1,2, Andreza Raquel Barbosa de Farias3, Gilberto Barbosa Domont2, Francisco de Assis de Paiva Campos4, Fábio César Sousa Nogueira5,6. 1. Laboratory of Proteomics/LADETEC, Universidade Federal Do Rio de Janeiro, Rio de Janeiro, Brazil. 2. Proteomics Unit, Institute of Chemistry, Universidade Federal Do Rio de Janeiro, Rio de Janeiro, Brazil. 3. Department of Agricultural Sciences, Universidade Federal Do Ceará, Fortaleza, 60356-900, Brazil. 4. Department of Biochemistry and Molecular Biology, Universidade Federal Do Ceará, Fortaleza, Brazil. bioplant@ufc.br. 5. Laboratory of Proteomics/LADETEC, Universidade Federal Do Rio de Janeiro, Rio de Janeiro, Brazil. fabiocsn@iq.ufrj.br. 6. Proteomics Unit, Institute of Chemistry, Universidade Federal Do Rio de Janeiro, Rio de Janeiro, Brazil. fabiocsn@iq.ufrj.br.
Abstract
BACKGROUND: Casbene synthase (CS) is responsible for the first committed step in the biosynthesis of phorbol esters (PE) in the Euphorbiaceae. PE are abundant in the seeds of the biofuel crop Jatropha curcas and its toxicity precludes the use of the protein-rich cake obtained after oil extraction as an animal feed and the toxicity of the fumes derived from burning PE containing biofuel is also a matter of concern. This toxicity is a major hindrance to exploit the potential of this crop as a source of raw material to produce biodiesel. For this reason, the current research on J. curcas is mainly focused on the understanding of the biosynthesis and site of synthesis of PE, as an avenue for the development of genotypes unable to synthesize PE in its seeds. RESULTS: Here, we present targeted proteomics assays (SRM and PRM) to detect and quantify CS in leaves, endosperm, and roots of two J. curcas genotypes with contrasting levels of PE. These assays were based on the use of reference isotopic labeled synthetic peptides (ILSP) predicted from 12 gene models of CS from the J. curcas genome. CONCLUSION: Our targeted proteomics methods were able to detect and quantify, for the first time, CS gene products and demonstrate the distribution of CS isoforms only in roots from J. curcas genotypes with a high and low concentration of PE. These methods can be expanded to monitor CS, at the protein level, in different tissues and genotypes of J. curcas.
BACKGROUND:Casbene synthase (CS) is responsible for the first committed step in the biosynthesis of phorbol esters (PE) in the Euphorbiaceae. PE are abundant in the seeds of the biofuel crop Jatropha curcas and its toxicity precludes the use of the protein-rich cake obtained after oil extraction as an animal feed and the toxicity of the fumes derived from burning PE containing biofuel is also a matter of concern. This toxicity is a major hindrance to exploit the potential of this crop as a source of raw material to produce biodiesel. For this reason, the current research on J. curcas is mainly focused on the understanding of the biosynthesis and site of synthesis of PE, as an avenue for the development of genotypes unable to synthesize PE in its seeds. RESULTS: Here, we present targeted proteomics assays (SRM and PRM) to detect and quantify CS in leaves, endosperm, and roots of two J. curcas genotypes with contrasting levels of PE. These assays were based on the use of reference isotopic labeled synthetic peptides (ILSP) predicted from 12 gene models of CS from the J. curcas genome. CONCLUSION: Our targeted proteomics methods were able to detect and quantify, for the first time, CS gene products and demonstrate the distribution of CS isoforms only in roots from J. curcas genotypes with a high and low concentration of PE. These methods can be expanded to monitor CS, at the protein level, in different tissues and genotypes of J. curcas.
Authors: Wei He; Andrew J King; M Awais Khan; Jesús A Cuevas; Danièle Ramiaramanana; Ian A Graham Journal: Plant Physiol Biochem Date: 2011-07-23 Impact factor: 4.270
Authors: Mohibullah Shah; Emanoella L Soares; Magda L B Lima; Camila B Pinheiro; Arlete A Soares; Gilberto B Domont; Fabio C S Nogueira; Francisco A P Campos Journal: J Proteomics Date: 2016-02-27 Impact factor: 4.044
Authors: Emanoella L Soares; Mohibullah Shah; Arlete A Soares; José H Costa; Paulo Carvalho; Gilberto B Domont; Fábio C S Nogueira; Francisco A P Campos Journal: J Proteome Res Date: 2014-07-22 Impact factor: 4.466