Open AccessPulsed EPR Distance Measurements in Soluble Proteins by Site‐Directed Spin Labeling (SDSL)
Author(s) -
Vera Ian Mitchelle S.,
Blackburn Mandy E.,
Galiano Luis,
Fanucci Gail E.
Publication year - 2013
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/0471140864.ps1717s74
Subject(s) - site directed spin labeling , electron paramagnetic resonance , spin label , chemistry , nuclear magnetic resonance , biophysics , physics , biology
The resurgence of pulsed electron paramagnetic resonance (EPR) in structural biology centers on recent improvements in distance measurements using the double electron‐electron resonance (DEER) technique. This unit focuses on EPR‐based distance measurements by site‐directed spin labeling (SDSL) of engineered cysteine residues in soluble proteins, with HIV‐1 protease used as a model. To elucidate conformational changes in proteins, experimental protocols were optimized and existing data analysis programs were employed to derive distance‐distribution profiles. Experimental considerations, sample preparation, and error analysis for artifact suppression are also outlined herein. Curr. Protoc. Protein Sci . 74:17.17.1‐17.17.29. © 2013 by John Wiley & Sons, Inc.