Saturation Recovery EPR and Nitroxide Spin Labeling for Exploring Structure and Dynamics in Proteins. Methods Enzymol 2015;564:3-27
Date
10/20/2015Pubmed ID
26477246DOI
10.1016/bs.mie.2015.07.016Scopus ID
2-s2.0-84944352221 (requires institutional sign-in at Scopus site) 19 CitationsAbstract
Experimental techniques capable of determining the structure and dynamics of proteins are continuously being developed in order to understand protein function. Among existing methods, site-directed spin labeling in combination with saturation recovery (SR) electron paramagnetic resonance spectroscopy contributes uniquely to the determination of secondary and tertiary protein structure under physiological conditions, independent of molecular weight and complexity. In addition, SR of spin labeled proteins was recently demonstrated to be sensitive to conformational exchange events with characteristic lifetimes on the order of μs, a time domain that presents a significant challenge to other spectroscopic techniques. In this chapter, we present the theoretical background necessary to understand the capabilities of SR as applied to spin labeled proteins, the instrumental requirements, and practical experimental considerations necessary to obtain interpretable data, and the use of SR to obtain information on protein: (1) secondary structure via solvent accessibility measurements, (2) tertiary structure using interspin distance measurements, and (3) conformational exchange.
Author List
Yang Z, Bridges M, Lerch MT, Altenbach C, Hubbell WLAuthor
Michael Lerch PhD Assistant Professor in the Biophysics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AlgorithmsAnimals
Electron Spin Resonance Spectroscopy
Humans
Models, Molecular
Protein Conformation
Proteins
Spin Labels
