Stage Master 2

Il y a 2 jours

Rennes, Bretagne, France Inserm Temps plein

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Internship in Chemical Biology

DNA scaffolds for tunable spatiotemporal control over receptor kinase assemblies.

Scientific background – Cellular stress induced by the abnormal accumulation of improperly folded proteins in the endoplasmic reticulum (ER) is emerging as a major actor in disease development and an appealing actionable target.[1] ER stress levels are under constant surveillance by the unfolded protein response (UPR), a major adaptive mechanism that lies at the core of cellular homeostasis and is responsible for cellular life-or-death decisions. The Inositol-requiring enzyme 1 (IRE1), the most evolutionary conserved UPR transducer, is an ER-resident transmembrane protein with a cytosolic dual kinase/RNase activity controlling pro-survival or pro-death signals. Yet, despite >20 years of investigations, the precise molecular mechanisms by which IRE1 is activated and exerts its catalytic and scaffolding functions still remain unclear and a subject of debate.[2] The group aims at deciphering the molecular mechanisms of IRE1 activation through a unique and novel prism at the interface of chemical biology, supramolecular chemistry and structural biology.

Description of the project & duties - This internship is in support of a PhD project focusing on using a programmable synthetic platform made of DNA structures (i.e. DNA origami) to control the nanoscale organization and nature of different complexes of a kinase protein.[3]

As part of this internship, you will focus on:

1 –Expressing, purifying and functionalizing the protein kinase N-terminus using a small molecule reagent.

2 - Assembling and purifying DNA origami structures.

3 - Bioconjugating proteins on the DNA origami using DNA hybridization principles.

4- Validating by AFM microscopy the functionalized Protein-DNA nanostructure obtained.

5 – Evaluating the enzymatic activities of the protein using established biochemical assays and molecular biology techniques.

Main activities and techniques involved –

• Molecular biology (design and conception of plasmids).

• Protein expression in eukaryotic cells.

• Protein engineering, bioconjugation, purification (affinity & SEC chromatography).

• Enzymatic assays.

• Structural characterization of protein-DNA conjugates (AFM, CryoEM).

References: [1] iScience 2023, 106687. https://doi.org/10.1016/j.isci.2023.106687; [2] Biomedicines 9, 156. https://doi.org/10.3390/biomedicines9020156; [3] Nat. Catal. 2020, 3, 295–306. https://doi.org/10.1038/s41929-019-0403-7.

Applicant profile – We encourage applications from highly motivated students from the national and international community with a background in chemical biology, biochemistry, biophysics, molecular biology, or other life sciences and with outstanding qualifications. Application only on this platform.

Stage Master 2

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Stage Master 2

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Emplois similaires à Rennes

Villes principales