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Biochemical and structural characterization of newly identified Uranium-binding proteins from plants



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Published on 25 March 2021
Project summary
Uranium (U) is a natural element that is mainly redistributed in the environment due to anthropogenic activities. Contamination of soils and water by U and its absorption by plants represent a significant health risk for human beings. The aim of the team project is to gain insight into the molecular mechanisms governing the fate of U in plants. This includes the characterization of U transporters as well as the identification of U-targeted cellular proteins.
The goal of the present student project is the biochemical and structural characterization of U-binding proteins from plants that we recently identified using metalloproteomic approaches (Immobilized U affinity chromatography and conventional chromatographic and electrophoretic techniques, coupled to ICP-MS for metal profiling, and tandem mass spectrometry for protein identification). These proteins can be either the cellular targets of toxicity or actors of detoxification mechanisms. Upon the 91 U-binding candidate proteins identified, 2 from the most promising ones will be further characterized in this project. These proteins were chosen because of their known capacity to bind metals and of their amino acid composition (U binds preferentially to acidic residues). Recombinant U-binding proteins will be produced in Escherichia coli, purified and their ability to bind U and other metals will be checked. Metal binding assays in vitro by ICP-MS and tryptophan fluorescence titration will be used to determine the stoichiometry of metal binding and the dissociation constant for the complexes, respectively. Depending on the nature of the identified targets the effect of uranyl binding will be performed by thermal stability and circular dichroism analyses (structural proteins) or activity measurements (enzymes). To gain insight into the mechanisms of U binding to targets, the 3D structures determination of U-protein complexes will be initiated by either X-ray crystallography (preferred approach), solution-state NMR spectroscopy (small proteins) or alternatively by cryo-electronic microscopy (large complexes).

Host laboratory
MetalStress team.

Keywords
uranium, protein, structure.

Supervisor
Claude Alban.


Call for students applications. Academic year 2021-2022

In the framework of the Chemistry Biology Health (CBH) Graduate School of Université Grenoble Alpes, the GRAL research programme funds MSc (Master 2) research scholarships.

We seek motivated candidates planning to:

attend one of the following Master programmes (2nd year MSc / Master 2 / M2) at Université Grenoble Alpes (taught in English):
Master in Biology:
Structural Biology of Pathogens
Immunology, Microbiology, Infectious Diseases
Neurobiology, Neurosciences
Physiology, Epigenetics Development, Cell Differentiation

Master in Plant Biology:
PLANTA International


& do a research internship in Integrated & Structural Cell Biology at one of the labs associated with the GRAL programme in Grenoble: IBS, PCV, BS, and CBM.

A maximum of 6 GRAL scholarships of 8 000 € are offered to cover the academic year from September 2021 to June 2022, which includes 4 months of courses and 6 months of research internship.

The call is open to EU and non-EU students.

The candidates must send their application in a unique pdf file to anne-mathilde.thierry _at_ ibs.fr (e-mail subject: M2-2021-LastName) before May 14th 2021 (23:59, CET), including:

A cover letter indicating the selected MSc programme at Université Grenoble Alpes, and 2 choices (ranked) for the 6-month research internship project
A CV
Diplomas and copies of academic transcripts for the last 2 academic years (translated to French or English if appropriate)
For non-native English speakers, level of language proficiency (e.g. TOEIC, Bulats, etc.)
One letter of recommendation
E-candidate portal application number

We strongly recommend that the candidates take contact with the potential supervisor(s) for the research project(s) of interest.

The applications will be evaluated by a selection committee composed of representatives of the GRAL Directory and the Master programmes.

The scholarship is conditional to the registration to one of the above programmes of the master’s degree in Biology of Université Grenoble Alpes, and to the recipient maintaining a satisfactory academic record in the programme. The admission to a 2nd year MSc programme (M2) at Université Grenoble Alpes is fully independent of the GRAL Scholarship. Application deadlines may differ between the different programmes. See admission requirements on Université Grenoble Alpes’s website: http://www.univ-grenoble-alpes.fr/fr/grandes-missions/formation/candidatures-et-inscriptions/


Schedule

Opening of the call for applications: March 1st 2021
Applications must be submitted by May 14th 2021
Results: June 2021

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