University of Minnesota
School of Physics & Astronomy

Biophysics Seminar

Thursday, October 5th 2017
10:10 am:
Biophysics Seminar in 120 PAN
Speaker: Ahmed Haikal, University of Minnesota, Duluth
Subject: Macromolecular Crowding Effects on Excited-State Dynamics of Hetero-FRET Probes in Mimetic Environments

Macromolecular crowding is prevalent in all living cells due to the presence of proteins, nucleic acids, and organelles. Cellular crowding is heterogeneous and is known to influence biomolecular transport, biochemical reactions, and protein assembly and folding. Emerging evidence suggests that some cell pathologies may be correlated with compartmentalized crowding. As a result, there is a need for robust biosensors to quantify crowding both spatially and temporally in living cells. Importantly, those biosensors should be compatible with noninvasive and quantitative micro-spectroscopy methods. I will highlight our recent wavelength-dependent excited-state dynamics of new hetero-FRET (mCerulean–linker–mCitrine) biosensors in environments of varying homogeneous and heterogeneous viscosities. Using time-resolved fluorescence and polarization anisotropy measurements, we elucidate the effects of the linker length and flexibility on the energy transfer efficiency as a function of the surrounding environment. Our results represent an important step towards the application of quantitative and non-invasive approaches to investigate macromolecular crowding and protein-protein interactions in living cells using hetero-FRET probes.

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