Title: Global responses to phenol and myrcene in Pseudomonas at the level of cell membrane proteome unveiled by quantitative proteomics.
Abstract: Mechanistic insights into Pseudomonas membrane proteome adaptation to phenol or myrcene were obtained during this thesis work by exploring quantitative proteomics.
Phenol-induced-alterations of the model-strain P. putida KT2440 membrane proteome were identified following total membranes purification and inner and outer membranes fractionation. Quantitative proteomic analysis was based on membrane proteins separation using two-dimensional gel electrophoresis (2-DE), their fluorescence staining, gel image analysis with Progenesis SameSpots software and protein identification by mass spectrometry. The adaptive response was found to include the coordinate increase of the content of solvent efflux pumps and the decreased content of porins, apparently also involving post-translational modifications (Roma-Rodrigues et al, 2010).
The modifications occurring in the membrane proteome of the biotechnologically-relevant Pseudomonas sp. M1 grown using myrcene as carbon source, compared with cells grown on lactate, were also unveiled based on isobaric-tags for relative and absolute quantitation (iTRAQ). This quantitative proteomic analysis revealed that myrcene leads to the alteration of cell surface towards increased hydrophobicity, and to the decreased content of electron transport chain proteins, possibly due to a switch to nitrate/nitrite respiration. Results also suggested a simultaneous use of lactate and myrcene as carbon sources and a possible association of some myrcene catabolic enzymes with cell membrane (Roma-Rodrigues et al, unpublished results).
Acknowledgments: This study was financially supported by “Fundação para a Ciência e a Tecnologia” (contracts PTDC/EBB-BIO/104980/2008 and PhD grant SFRH/BD/38805/2007).
Roma-Rodrigues C, Santos PM, Benndorf D, Rap E, Sá-Correia, I. (2010), Response of Pseudomonas putida KT2440 to phenol at the level of membrane proteome. J. Proteomics 73: 1461-1478.
PhD Degree: Biotechnology; Year of conlusion: 2012
Institution: Instituto Superior Técnico, Universidade Técnica de Lisboa
Supervision: Isabel Sá-Correia (IST/UTL) and Pedro Miguel Santos (UM)