Towards a realistic model of the cellular environment: Dynamics in 'naturally' crowded protein solutions
Using deuterated cellular lysate obtained from living cells, we will establish a natural crowding environment for protonated "tracer" or "target" immunoglobulin (Ig) proteins. In a previous experiment, we were able to identify protein diffusion and an internal dynamical process in the QENS signal of IN16B in standard configuration. Given the rather low Ig concentrations and the presence of lysate, IN16B in the standard configuration probes mainly the global diffusion and the slow part of the internal dynamics. To obtain a comprehensive picture of both the slow and fast internal dynamics in such a "naturally" crowded environment, a broader dynamic range is needed. We plan to systematically investigate the influence of the presence of lysate on the internal diffusive motions of the target proteins using BATS. In this way, we will maintain a good overlap with existing IN16B data, while having a dynamic range sufficiently large to better determine the antibody fast internal dynamics. The study will be also an essential methodological step to establish a reliable access to protein dynamics under native-like crowding conditions.
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GRIMALDO Marco; APPEL Markus; BECK Christian; DA VELA Stefano; Juliette M. Devos; FORSYTH Victor Trevor; HAERTLEIN Michael; LAUX Valerie; MATSARSKAIA Olga; MOULIN Martine; ROOSEN RUNGE Felix; Frank Schreiber; SEYDEL Tilo and ZHANG Fajun. (2019). Towards a realistic model of the cellular environment: Dynamics in 'naturally' crowded protein solutions. Institut Laue-Langevin (ILL) doi:10.5291/ILL-DATA.8-04-855
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