Berre-les-Alpes, France
Berre-les-Alpes, France

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Trauchessec M.,CEA Grenoble | Trauchessec M.,French Institute of Health and Medical Research | Trauchessec M.,UniversiteGrenoble Alpes | Jaquinod M.,CEA Grenoble | And 20 more authors.
Molecular and Cellular Proteomics | Year: 2014

Metabolic engineering aims to design high performance microbial strains producing compounds of interest. This requires systems-level understanding; genome-scale models have therefore been developed to predict metabolic fluxes. However, multi-omics data including genomics, transcriptomics, fluxomics, and proteomics may be required to model the metabolism of potential cell factories. Recent technological advances to quantitative proteomics have made mass spectrometry-based quantitative assays an interesting alternative to more traditional immuno- affinity based approaches. This has improved specificity and multiplexing capabilities. In this study, we developed a quantification workflow to analyze enzymes involved in central metabolism in Escherichia coli (E. coli). This workflow combined full-length isotopically labeled standards with selected reaction monitoring analysis. First, full-length 15N labeled standards were produced and calibrated to ensure accurate measurements. Liquid chromatography conditions were then optimized for reproducibility and multiplexing capabilities over a single 30-min liquid chromatography-MS analysis. This workflow was used to accurately quantify 22 enzymes involved in E. coli central metabolism in a wild-type reference strain and two derived strains, optimized for higher NADPH production. In combination with measurements of metabolic fluxes, proteomics data can be used to assess different levels of regulation, in particular enzyme abundance and catalytic rate. This provides information that can be used to design specific strains used in biotechnology. In addition, accurate measurement of absolute enzyme concentrations is key to the development of predictive kinetic models in the context of metabolic engineering. © 2014 by The American Society for Biochemistry and Molecular Biology.


Thalmann C.,ETH Zurich | Mulders G.D.,University of Arizona | Janson M.,University of Stockholm | Olofsson J.,Max Planck Institute for Astronomy | And 17 more authors.
Astrophysical Journal Letters | Year: 2015

We present the first optical (590-890 nm) imaging polarimetry observations of the pre-transitional protoplanetary disk around the young solar analog LkCa 15, addressing a number of open questions raised by previous studies. We detect the previously unseen far side of the disk gap, confirming the highly off-centered scattered-light gap shape that was postulated from near-infrared imaging, at odds with the symmetric gap inferred from millimeter interferometry. Furthermore, we resolve the inner disk for the first time and trace it out to 30 AU. This new source of scattered light may contribute to the near-infrared interferometric signal attributed to the protoplanet candidate LkCa 15 b, which lies embedded in the outer regions of the inner disk. Finally, we present a new model for the system architecture of LkCa 15 that ties these new findings together. These observations were taken during science verification of SPHERE ZIMPOL and demonstrate this facility's performance for faint guide stars under adverse observing conditions. © 2015. The American Astronomical Society. All rights reserved.

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