Anne-Gaëlle Planson

Microbiologie de l’alimentation au service de la santé

Author Statistics

Papers

Citation

H-Index

i-10 index

Research Trends

Author Order

Document Type

Co-Authors

Jean‐Loup Faulon

Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement

Pablo Carbonell

Universitat Politècnica de València

Matthieu Jules

Microbiologie de l’alimentation au service de la santé

Michel B. Tolédano

Mayo Clinic

Alain Chaffotte

Laboratoire des Biomolécules

Ioana Grigoras

Centre National de la Recherche Scientifique

A Delaunay

Institut de Biologie Intégrative de la Cellule

Etienne Dervyn

Microbiologie de l’alimentation au service de la santé

Jon Beckwith

Harvard University

Jacqueline Cherfils

Centre National de la Recherche Scientifique

Cooperative Institutions

Centre National de la Recherche Scientifique

Université Paris-Saclay

Commissariat à l'Énergie Atomique et aux Énergies Alternatives

Inserm

CEA Paris-Saclay

Université Paris-Sud

Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement

Laboratoire d'Enzymologie et Biochimie Structurales

Université Paris Cité

Harvard University

Author Statistics

Papers

Citation

H-Index

i-10 index

Research Field

Inhibitory signalling to the Arp2/3 complex steers cell migration

Nature (2013)

Irène Dang Roman Gorelik Carla Sousa-Blin Emmanuel Derivery Christophe Guérin

Lamellipodium

Pseudopodia

10.1038/nature12611

Cite

Citations (233)

Validation of RetroPath, a computer‐aided design tool for metabolic pathway engineering

Biotechnology Journal (2014)

Tamás Fehér Anne-Gaëlle Planson Pablo Carbonell Alfred Fernández‐Castané Ioana Grigoras

Metabolic engineering has succeeded in biosynthesis of numerous commodity or high value compounds. However, the choice of pathways and enzymes used for production was many times made ad hoc, or required expert knowledge of the specific biochemical reactions. In order to rationalize the process of engineering producer strains, we developed the computer-aided design (CAD) tool RetroPath that explores and enumerates metabolic pathways connecting the endogenous metabolites of a chassis cell to the target compound. To experimentally validate our tool, we constructed 12 top-ranked enzyme combinations producing the flavonoid pinocembrin, four of which displayed significant yields. Namely, our tool queried the enzymes found in metabolic databases based on their annotated and predicted activities. Next, it ranked pathways based on the predicted efficiency of the available enzymes, the toxicity of the intermediate metabolites and the calculated maximum product flux. To implement the top-ranking pathway, our procedure narrowed down a list of nine million possible enzyme combinations to 12, a number easily assembled and tested. One round of metabolic network optimization based on RetroPath output further increased pinocembrin titers 17-fold. In total, 12 out of the 13 enzymes tested in this work displayed a relative performance that was in accordance with its predicted score. These results validate the ranking function of our CAD tool, and open the way to its utilization in the biosynthesis of novel compounds.

Metabolic Engineering

Metabolic network

Metabolic pathway

Pinocembrin

Synthetic Biology

10.1002/biot.201400055

Cite

Citations (55)

Determinants of activity in glutaredoxins: an in vitro evolved Grx1-like variant of Escherichia coli Grx3

Biochemical Journal (2010)

Tobias H. Elgán Anne-Gaëlle Planson Jon Beckwith Peter Güntert Kurt D. Berndt

The Escherichia coli glutaredoxins 1 and 3 (Grx1 and Grx3) are structurally similar (37% sequence identity), yet have different activities in vivo. Unlike Grx3, Grx1 efficiently reduces protein disulfides in proteins such as RR (ribonucleotide reductase), whereas it is poor at reducing S-glutathionylated proteins. An E. coli strain lacking genes encoding thioredoxins 1 and 2 and Grx1 is not viable on either rich or minimal medium; however, a M43V mutation in Grx3 restores growth under these conditions and results in a Grx1-like protein [Ortenberg, Gon, Porat and Beckwith (2004) Proc. Natl. Acad. Sci. U.S.A. 101, 7439-7944]. To uncover the structural basis of this change in activity, we have compared wild-type and mutant Grx3 using CD and NMR spectroscopy. Ligand-induced stability measurements demonstrate that the Grx3(M43V/C65Y) mutant has acquired affinity for RR. Far-UV CD spectra reveal no significant differences, but differences are observed in the near-UV region indicative of tertiary structural changes. NMR (1)H-(15)N HSQC (heteronuclear single quantum coherence) spectra show that approximately half of the 82 residues experience significant (Deltadelta>0.03 p.p.m.) chemical shift deviations in the mutant, including nine residues experiencing extensive (Deltadelta > or =0.15 p.p.m.) deviations. To test whether the M43V mutation alters dynamic properties of Grx3, H/D (hydrogen/deuterium) exchange experiments were performed demonstrating that the rate at which backbone amides exchange protons with the solvent is dramatically enhanced in the mutant, particularly in the core of the protein. These data suggest that the Grx1-like activity of the Grx3(M43V/C65Y) mutant may be explained by enhanced intrinsic motion allowing for increased specificity towards larger substrates such as RR.

Glutaredoxin

Ribonucleotide reductase

10.1042/bj20100289

Cite

Citations (9)

Reining in H2O2 for Safe Signaling

Cell (2010)

Michel B. Tolédano Anne-Gaëlle Planson A Delaunay

10.1016/j.cell.2010.02.003

Cite

Citations (50)

Sulfiredoxin Protects Mice from Lipopolysaccharide-Induced Endotoxic Shock

Antioxidants and Redox Signaling (2010)

Anne-Gaëlle Planson Gaël Palais Kahina Abbas Matthieu Gérard Linhdavanh Couvelard

Peroxiredoxins constitute a major family of cysteine-based peroxide-scavenging enzymes. They carry an intriguing redox switch by undergoing substrate-mediated inactivation via overoxidation of their catalytic cysteine to the sulfinic acid form that is reverted by reduction catalyzed by the sulfinic acid reductase sulfiredoxin (Srx). The biological significance of such inactivation is not understood, nor is the function of Srx1. To address this question, we generated a mouse line with a null deletion of the Srx1-encoding Srxn1 gene. We show here that Srxn1(-/-) mice are perfectly viable and do not suffer from any apparent defects under laboratory conditions, but have an abnormal response to lipopolysaccharide that manifests by increased mortality during endotoxic shock. Microarray-based mRNA profiles show that although the response of Srxn1(-/-) mice to lipopolysaccharide is typical, spanning all spectrum and all pathways of innate immunity, it is delayed by several hours and remains intense when the response of Srxn1(+/+) mice has already dissipated. These data indicate that Srx1 activity protects mice from the lethality of endotoxic shock, adding this enzyme to other host factors, as NRF2 and peroxiredoxin 2, which by regulating cellular reactive oxygen species levels act as important modifiers in the pathogenesis of sepsis.

Peroxiredoxin

10.1089/ars.2010.3552

Cite

Citations (32)

A retrosynthetic biology approach to metabolic pathway design for therapeutic production

BMC Systems Biology (2011)

Pablo Carbonell Anne-Gaëlle Planson Davide Fichera Jean‐Loup Faulon

Abstract Background Synthetic biology is used to develop cell factories for production of chemicals by constructively importing heterologous pathways into industrial microorganisms. In this work we present a retrosynthetic approach to the production of therapeutics with the goal of developing an in situ drug delivery device in host cells. Retrosynthesis, a concept originally proposed for synthetic chemistry, iteratively applies reversed chemical transformations (reversed enzyme-catalyzed reactions in the metabolic space) starting from a target product to reach precursors that are endogenous to the chassis. So far, a wider adoption of retrosynthesis into the manufacturing pipeline has been hindered by the complexity of enumerating all feasible biosynthetic pathways for a given compound. Results In our method, we efficiently address the complexity problem by coding substrates, products and reactions into molecular signatures. Metabolic maps are represented using hypergraphs and the complexity is controlled by varying the specificity of the molecular signature. Furthermore, our method enables candidate pathways to be ranked to determine which ones are best to engineer. The proposed ranking function can integrate data from different sources such as host compatibility for inserted genes, the estimation of steady-state fluxes from the genome-wide reconstruction of the organism's metabolism, or the estimation of metabolite toxicity from experimental assays. We use several machine-learning tools in order to estimate enzyme activity and reaction efficiency at each step of the identified pathways. Examples of production in bacteria and yeast for two antibiotics and for one antitumor agent, as well as for several essential metabolites are outlined. Conclusions We present here a unified framework that integrates diverse techniques involved in the design of heterologous biosynthetic pathways through a retrosynthetic approach in the reaction signature space. Our engineering methodology enables the flexible design of industrial microorganisms for the efficient on-demand production of chemical compounds with therapeutic applications.

Retrosynthetic analysis

Synthetic Biology

Metabolic Engineering

Metabolic pathway

10.1186/1752-0509-5-122

Cite

Citations (108)

Assistance of Maltose Binding Protein to the in Vivo Folding of the Disulfide-Rich C-Terminal Fragment from Plasmodium falciparum Merozoite Surface Protein 1 Expressed in Escherichia coli

Biochemistry (2003)

Anne-Gaëlle Planson J. Iñaki Guijarro Michel Goldberg Alain Chaffotte

The C-terminal fragment of Plasmodium falciparum merozoite surface protein 1 (F19) is a leading candidate for the development of a malaria vaccine. Successful vaccination trials on primates, immunochemistry, and structural studies have shown the importance of its native conformation for its protective role against infection. F19 is a disulfide-rich protein, and the correct pairing of its 12 half-cystines is required for the native state of the protein. F19 has been produced in the Escherichia coli periplasm, which has an oxidative environment favorable for the formation of disulfide bonds. F19 was either expressed as a fusion with the maltose binding protein (MBP) or directly addressed to the periplasm by fusing it with the MBP signal peptide. Direct expression of F19 in the periplasm led to a misfolded protein with a heterogeneous distribution of disulfide bridges. On the contrary, when produced as a fusion protein with E. coli MBP, the F19 moiety was natively folded. Indeed, after proteolysis of the fusion protein, the resulting F19 possesses the structural characteristics and the immunochemical reactivity of the analogous fragment produced either in baculovirus-infected insect cells or in yeast. These results demonstrate that the positive effect of MBP in assisting the folding of passenger proteins extends to the correct formation of disulfide bridges in vivo. Although proteins or protein fragments fused to MBP have been frequently expressed with success, our comparative study evidences for the first time the helping property of MBP in the oxidative folding of a disulfide-rich protein.

Maltose-binding protein

Oxidative folding

Proteolysis

10.1021/bi035321c

Cite

Citations (22)

New Insights for Native Production of MSP119, the Disulfide-Rich C-Terminal Fragment from Plasmodium falciparum Merozoite Surface Protein 1

PLoS ONE (2013)

Anne-Gaëlle Planson J. Iñaki Guijarro Alain Chaffotte

Malaria represents a major public health problem and an important cause of mortality and morbidity. The malaria parasites are becoming resistant to drugs used to treat the disease and still no efficient vaccine has been developed. One promising vaccine candidate is the merozoite surface protein 1 (MSP1), which has been extensively investigated as a vaccine target. The surface protein MSP1 plays an essential role in the erythrocyte invasion process and is an accessible target for the immune system. Antibodies to the carboxy-terminal region of the protein, named MSP119, can inhibit erythrocyte invasion and parasite growth. In order to develop an effective MSP119- based vaccine against malaria, production of an antigen that is recognized by protective antibodies is mandatory. To this aim, we propose a method to produce the disulfide-rich MSP119 in its native conformation based on its in vitro oxidative refolding. The native conformation of the renatured MSP119 is carefully established by immunochemical reactivity experiments, circular dichroism and NMR. MSP119 can successfully be refolded in vitro as an isolated protein or as a fusion with the maltose binding protein. The possibility to properly fold MSP119in vitro paves the way to new approaches for high titer production of native MSP119 using Escherichia coli as a host.

Maltose-binding protein

10.1371/journal.pone.0057086

Cite

Citations (4)

A retrosynthetic biology approach to therapeutics: from conception to delivery

Current Opinion in Biotechnology (2012)

Anne-Gaëlle Planson Pablo Carbonell Ioana Grigoras Jean‐Loup Faulon

Retrosynthetic analysis

Synthetic Biology

10.1016/j.copbio.2012.03.009

Cite

Citations (22)