Proteome of the phytopathogen Xanthomonas citri subsp. citri: a global expression profile

Abstract

BackgroundCitrus canker is a disease caused by Xantomonas citri subsp.citri (Xac), and has emerged as one of the major threats to the worldwide citrus crop because it affects all commercial citrus varieties, decreases the production and quality of the fruits and can spread rapidly in citrus growing areas. In this work, the first proteome of Xac was analyzed using two methodologies, two-dimensional liquid chromatography (2D LC) and tandem mass spectrometry (MS/MS).ResultsIn order to gain insight into the metabolism of Xac, cells were grown on two different media (NB - Nutrient Broth and TSE - Tryptone Sucrose broth enriched with glutamic acid), and proteins were proteolyzed with trypsin and examined by 2D LC-MS/MS. Approximately 39% of all predicted proteins by annotation of Xac were identified with their component peptides unambiguously assigned to tandem mass spectra. The proteins, about 1,100, were distributed in all annotated functional categories.ConclusionsThis is the first proteomic reference map for the most aggressive strain of Xanthomonas pathogen of all orange varieties. The compilation of metabolic pathways involved with bacterial growth showed that Xac expresses a complete central and intermediary metabolism, replication, transcription and translation machineries and regulation factors, distinct membrane transporters (ABC, MFS and pumps) and receptors (MCP, TonB dependent and metabolites acquisition), two-component systems (sensor and regulatory components) and response regulators. These data corroborate the growth curve in vitro and are the first reports indicating that many of these genome annotated genes are translated into operative in Xac. This proteomic analysis also provided information regarding the influence of culture medium on growth and protein expression of Xac.

Proteomic analysis of the reproductive tract fluids from tropically-adapted Santa Ines rams

“…These proteins are secreted in response to cellular damage and protect cells by preventing accumulation of misfolded proteins and blocking apoptotic pathways [62]. Clusterin was Protein data were analyzed using the software for researching annotations of proteins (STRAP [19]). Gene ontology terms were obtained from UniProtKB and EBI GOA databases.…”

“…Protein identification by electrospray ionization-quadrupole-time of flight (ESI-Q-ToF) mass spectrometry: spots from the seminal plasma, cauda epididymal and seminal vesicle fluids For LC-MS/MS [19], the digested samples were injected using the nanoAcquity UPLC sample manager and the chromatographic separation was performed using a UPLC C18 column (75 μm × 10 cm) with a flow of 0.6 μL/min. The mass spectra were acquired in a Synapt G2 HDMS instrument (Waters Co., Milford, MA, USA) using a data dependent acquisition (DDA), where the three top peaks were subjected to MS/MS.…”

Selection of a protein solubilization method suitable for phytopathogenic bacteria: a proteomics approach

“…Proteomic studies conducted with the bacterial phytopathogen Xanthomonas axonopodis pv. citri showed that the Phenol method was employed with success [26]. In some cases, it is necessary to develop a new method due to peculiarities of the sample in question as noted by Barbarino and Lourenço in 2005 [27] due to high concentrations of salts present in the sample.…”

A proteome map of primary cultured rat Schwann cells

“…The gel-based proteomics strategies are being replaced rapidly by a new proteomics strategy based on liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) [5-8], which shows a high degree of specificity and sensitivity.…”