Dataset Information

Monitoring Casbene Synthase in Jatropha curcas Tissues Using Targeted Proteomics

ABSTRACT: Casbene synthase is responsible for the first commited 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 for the production of biodiesel. For this reason, current research on J. curcas is mainly focused in the understanding of the biosynthesis and site of synthesis of PE, as an avenue for the development of biotechnological of genotypes unable to synthesize PE in its seeds. Here, we present the results of targeted proteome assays (SRM and PRM) to monitor and quantify casbene synthase in leaves, endosperma and roots of two J. curcas of contrasting levels of PE. The assays were based in the use of synthetic isotopic labeled peptides derived from 12 gene models of casbene synthase from the J. curcas genome and results showed the presence of casbene synthase encoded in seven of the 12 gene models. Several specific transitions were identified and which can be used to monitor several casbene synthase proteins by any one of the two targeted proteomics assays and/or to validate the results of transcription-based experiments.

INSTRUMENT(S): TSQ Quantiva, Q Exactive Plus

ORGANISM(S): Jatropha Curcas

TISSUE(S): Root, Endosperm, Leaf

SUBMITTER: Proteomics Unit

LAB HEAD: Fábio César Sousa Nogueira

PROVIDER: PXD019525 | Pride | 2021-09-09

REPOSITORIES: Pride

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Publications

Monitoring casbene synthase in Jatropha curcas tissues using targeted proteomics.

de Almeida Natália Pinto NP Neto Domingos Ferreira Mélo DFM Carneiro Gabriel Reis Alves GRA de Farias Andreza Raquel Barbosa ARB Domont Gilberto Barbosa GB de Paiva Campos Francisco de Assis FA Nogueira Fábio César Sousa FCS

Plant methods 20210206 1

<h4>Background</h4>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 ...[more]

PMID: 33549129

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Project description:Phase 1: The whole saliva samples were obtained from 66 systemically healthy subjects consisting of patients with chronic periodontitis (CP), generalized aggressive periodontitis (G-AgP), gingivitis and individuals with periodontal health. The use of humans for study satisfied the requirements of the Ege University Institutional Review Board (Ethics number 16-12.1/16) and and was conducted in accordance with the guidelines of the World Medical Association Declaration of Helsinki. It is confirmed that this cross-sectional study conforms to STROBE guidelines for observational studies. Complete medical and dental histories were taken from all participants. Systemic exclusion criteria were the presence of cardiovascular and respiratory diseases, diabetes mellitus, HIV infection, systemic inflammatory conditions or non-plaque-induced oral inflammatory conditions, immunosuppressive chemotherapy, and current pregnancy or lactation. Smoking status was also recorded. None of the patients had taken medication such as antibiotics or anti-inflammatory drugs that could affect their periodontal status for at least 6 months before the study. Patients eligible for the study returned to the clinic for clinical measurement screening one-week after being pre-screened. Before being enrolled in the study, participants provided written and informed consent to use their saliva samples and individual data for scientific purposes. The clinical periodontal indices including probing depth (PPD), clinical attachment level (CAL), plaque index (PI) and bleeding on probing (BOP) were recorded by a manual periodontal probe by a trained and calibrated examiner (V.O.O). The extent and severity of alveolar bone support were evaluated radiographically in each patient. The participants were classified into four groups based on their periodontal conditions according to the criteria proposed by the 1999 International Workshop for a Classification of Periodontal Diseases and Conditions (REF). Generalized Aggressive Periodontitis (G-AgP): The G-AgP group included 17 patients with ≥16 teeth. The patients had a non- contributory medical history and demonstrated with an early age of clinical manifestations with a generalized pattern of rapid attachment loss and bone destruction. These individuals had minimum of CAL ≥ 5 mm and PD ≥ 6 mm on eight or more teeth; at least three of these were other than central incisors or first molars. Radiographic bone loss was above ≥ 30 % of root length affecting ≥3 permanent teeth other than first molars and incisors. Generalized Chronic Periodontitis (CP): The CP group included 17 patients with severe generalized chronic periodontitis. These individuals had a minimum four non-adjacent teeth with sites with CAL ≥ 5 mm and PPD ≥ 6 mm, and ≥50 % alveolar bone loss in at least two quadrants which was commensurate with the amount of plaque accumulation. Phase 2: We aimed to develop SRM-based workflow for detecting and quantitating the relative abundance of the candidate proteins in an independent saliva cohort . 82 subjects (48 females and 34 males; age range 24-59 years) were recruited for the study at the Department of Periodontology, School of Dentistry, Adnan Menderes University, Aydın, Turkey. Ethical clearance was obtained from the Ethics Committee of the School of Medicine, Ege University with the protocol number ((Ethics number 70198063-050.06.04). Each participant gave written and verbal informed consent after the purpose and procedures of the study were explained. A dental and medical history was compiled for all subjects as detailed in phase 1 and participants were clustered into four groups according to the criteria proposed by the 1999 International Workshop for a Classification of Periodontal Diseases and Conditions as detailed at Phase 1

Dataset Information

Monitoring Casbene Synthase in Jatropha curcas Tissues Using Targeted Proteomics

Publications

Monitoring casbene synthase in Jatropha curcas tissues using targeted proteomics.

Similar Datasets

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