Photo by nci from unsplash
Macromolecular crowding influences many intracellular processes such as protein folding, biochemical reactions and diffusion. Recently, a rationally designed FRET probe was developed to quantify macromolecular crowding in vitro (Boersma et… Click to show full abstract
Macromolecular crowding influences many intracellular processes such as protein folding, biochemical reactions and diffusion. Recently, a rationally designed FRET probe was developed to quantify macromolecular crowding in vitro (Boersma et al. Nat. Meth. 12:227, 2015). The probe consists of a FRET pair (mCerulean and mCitrine) that is connected with a flexible linker. Here, we characterized the same FRET probe in crowded solutions using fluorescence lifetime measurements as a function of the concentration of a crowding agent (Ficoll-70) as well as the excitation/detection wavelengths towards selective monitoring of the donor and acceptor.
Journal Title: Biophysical Journal
Year Published: 2017
Link to full text (if available)
Share on Social Media: Sign Up to like & get
recommendations!