@inbook{883a018d6bba4bd2b0a1fcc01f67894d,
title = "Short-Range Distance Measurement by Transition Metal Ion FRET",
abstract = "Measurement of atomic-scale conformational dynamics in proteins has proved a challenging endeavor, although these movements are pivotal for understanding the mechanisms behind protein function. Herein we describe a fluorescence-based method that enables the measurement of distances between specific domains within a protein and how it might change during protein function. The method is transition metal ion F{\"o}rster resonance energy transfer (tmFRET) and builds on the principle that the fluorescence emission from a fluorophore can be quenched in a distance-dependent manner by a colored transition metal such as nickel (Ni2+), copper (Cu2+), or cobalt (Co2+). It can be applied to literally any protein where it is possible to perform site-specific incorporation of a fluorescent molecule. This chapter will explain the use and applications of tmFRET in detail using incorporation of the dye with cysteine chemistry on a purified protein sample.",
keywords = "Conformational dynamics, Cysteine chemistry, Fluorescence spectroscopy, F{\"o}rster resonance energy transfer, Intramolecular distance measurements, Protein purification, Transition metals",
author = "Mortensen, {Jonas S.} and Loland, {Claus J.}",
year = "2020",
doi = "10.1007/978-1-0716-0724-4_14",
language = "English",
series = "Methods in Molecular Biology",
publisher = "Humana Press",
pages = "299--311",
booktitle = "Biophysics of membrane proteins",
address = "United States",
}