18 months Post-doctoral position at Univ. Grenoble Alpes/CEA, France on DNP-enhanced solid-state NMR for in-cell applications

29 Feb 2024
Job Information

Organisation/Company

Univ. Grenoble Alpes

Department

Labex Arcane

Research Field

Chemistry » Instrumental techniques

Researcher Profile

Recognised Researcher (R2)

Country

France

Application Deadline

31 May 2024 - 17:00 (Europe/Paris)

Type of Contract

Temporary

Job Status

Full-time

Offer Starting Date

1 Sep 2024

Is the job funded through the EU Research Framework Programme?

Not funded by an EU programme

Is the Job related to staff position within a Research Infrastructure?

No

Offer Description

Cellular applications of solid-state NMR are associated with major challenges: low sensitivity and resolution, need for isotopic labelling strategies able to suppress the large cellular background. Dynamic Nuclear Polarization (DNP) has emerged as a viable route to significantly improve the sensitivity of solid-state NMR, including for biomolecular and in-cell applications. In addition, DNP experiments are performed at low temperature (about 100 K or lower), which offers the additional advantage to protect the cell integrity. Some groups have started to investigate the potential of the technique towards in-cell applications, but the various approaches so far suffer from a lack of selectivity to detect low protein amounts under the large background signal of the cell, or required a highly deuterated cellular environment. In addition, spectral resolution is impaired by conformational heterogeneity induced by sample freezing.

We propose to solve the problems of resolution, sensitivity and selectivity associated with in-cell DNP experiments by developing an innovative approach based on the combination of bio-orthogonal chemistry and selective DNP (SelDNP). The methodology will be applied on mammalian cells for the study of Atox1, a copper (Cu) chaperone involved in Cu homeostasis.

Based on preliminary results, the methodology will be developed in stages, first on purified proteins, and then on the protein in mammalian cells. The first step will be to optimize experimental conditions for selective DNP (SelDNP) on Atox1 and to determine the best targeting strategy (including polarizing agent) that allows detecting sub-micromolar concentrations in cell lysates. This knowledge will then be used to target and detect Atox1 directly in the mammalian cell, using bio-orthogonal chemistry to introduce the spin label for SelDNP. SelDNP is a methodology developed over the last few years in our group. It enables the production of high-resolution DNP-enhanced NMR subspectra specific to a predefined region of the protein, determined by the position of the spin label. Demonstrated so far only on purified proteins, the methodology is very promising for targeting a biomolecule in the crowded environment of the cell, but as well for removing the cell background signal, which become significant at low protein concentrations.

Requirements

Research Field

Chemistry » Instrumental techniques

Education Level

PhD or equivalent

Skills/Qualifications

Applicants are expected to have a doctoral degree in liquid-state and/or solid-state biomolecular NMR spectroscopy. Knowledge about MAS-DNP will be considered as a plus. The candidate will mainly work in NMR and DNP, but depending on his/her expertise in biochemistry, he/she may assist in protein production and labeling.

Languages

ENGLISH

Level

Excellent

Additional Information

Website for additional job details

https://arcane.univ-grenoble-alpes.fr/opportunities-calls/job-offers

Work Location(s)

Number of offers available

1

Company/Institute

CEA / Interdisciplinary Research Institute of Grenoble

Country

France

City

Grenoble

Postal Code

38054

Street

17 rue des Martyrs

Geofield

Where to apply

E-mail

[email protected]

Contact

City

Grenoble

Website

https://nmr-dnp-grenoble.net/home/
https://arcane.univ-grenoble-alpes.fr/

STATUS: EXPIRED