Manjeet | Arcane - Université Grenoble Alpes

Methodological development for protein-selective DNP-enhanced NMR in mammalian cells

Cellular applications of solid-state nuclear magnetic resonance (NMR) are associated with major challenges: low sensitivity and resolution, requirement of isotopic labelling strategies 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. Most approaches proposed so far for in-cell DNP NMR 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. 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 SelDNP approach recently developed in the group enables the production of high-resolution DNP-enhanced NMR sub-spectra specific to a predefined region of the protein. 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. The methodology will be applied on mammalian cells for the study of Atox1, a copper (Cu) chaperone involved in Cu homeostasis.

My name is Manjeet. I received my PhD degree from Indian Institute of Technology Delhi. I am currently a postdoc fellow in the Modélisation et Exploration des Matériaux (MEM) and Laboratoire Chimie et Biologie des Métaux (LCBM) research units of Interdisciplinary Research Institute of Grenoble with Dr. Sabine Hediger (MEM) and Dr. Aurelien Deniaud (LCBM).

My area of research is nuclear magnetic resonance (NMR), an analytical tool that finds applications in different branches of science. During my PhD, I simply explored a few simple concepts to develop new simple methods. Up to now, I have not achieved anything great in my research career, nor I intend to. My simple desire to gain knowledge brought me on the path of research in science. This subconsciously motivated me to choose research as a career path and, now I intend to become an independent researcher.

Nuclear magnetic resonance has evolved, since its discovery, into an indispensable analytical tool in chemistry, biology, drug discovery, medicine, and so on. Given this diversity of NMR, I seek to diversify my skills in NMR by learning its different forms. I believe that I could leverage these skills to secure a research position. The work in my postdoctoral project involves different aspects of NMR---hence a perfect opportunity for me to learn these concepts. After the successful completion of this project, I will be looking for the job of an independent researcher.

I was unaware of the Labex Arcane program before applying for this postdoctoral position. However, I am now well aware of this program’s two scientific axes and its consortium of scientists from the University of Grenoble Alpes brought together to achieve its bio-driven goals.