abstract
Luminescent nanothermometry emerges as a powerful tool for studying protein dynamics. This technique was employed to perform the first measurement of the temperature dependence of protein Brownian velocity, showcasing the illustrative example of enhanced green fluorescent protein (EGFP) across physiologically relevant temperatures (30-50 degrees C) and concentrations (40, 60, and 80 x 10-3 kg m-3). EGFP exhibited a concentration-dependent decrease in Brownian velocity, from (1.47 +/- 0.09) x 10-3 m s-1 to (0.35 +/- 0.01) x 10-3 m s-1, at 30 degrees C, mimicking crowded cellular environments. Notably, the protein Brownian velocity increased linearly with temperature. These results demonstrate the suitability of concentrated suspensions for modeling intracellular crowding and validate luminescent nanothermometry for protein Brownian motion studies. Furthermore, the observed linear relationship between the logarithm of the protein Brownian velocity and concentration indicates that EGFP motion is not primarily driven by diffusion, but more of a ballistic transport. This study explores Brownian motion in enhanced green fluorescent protein (EGFP) using luminescence nanothermometry. How protein concentration and temperature impact EGFP dynamics is demonstrated providing insights into protein behavior in crowded environments. The research emphasizes the need to measure protein Brownian velocity at physiological temperatures for accurate biological studies. image
keywords
INSTANTANEOUS VELOCITY; DYNAMICS; LIQUIDS; MEMORY
subject category
Physics
authors
Guo, YW; Maturi, FE; Brites, CDS; Carlos, LD
our authors
Carlos Brites
Associate Professor
Fernando Eduardo Maturi
Research Fellowship
Guo Yongwei
PhD Student
Luís Carlos
Full professorProjects
CICECO - Aveiro Institute of Materials (UIDB/50011/2020)
CICECO - Aveiro Institute of Materials (UIDP/50011/2020)
Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)
acknowledgements
Y.G. and F.E.M. contributed equally to this work. This work was partially developed within the scope of the project CICECO-Aveiro Institute of Materials, UIDB/50011/2020 (DOI 10.54499/UIDB/50011/2020), UIDP/50011/2020 (DOI 10.54499/UIDP/50011/2020) & LA/P/0006/2020 (DOI 10.54499/LA/P/0006/2020), financed by national funds through the FCT/MCTES (PIDDAC). This work was also financially supported by the project Shape of Water (PTDC/NAN-PRO/3881/2020) through Portuguese funds. F.E.M. acknowledges the financial support from the FCT through Research Grant UI/BD/151445/2021 (DOI 10.54499/UI/BD/151445/2021). Y. G. acknowledges the financial support from the China Scholarship Council (No. 202306650010).