title: Pulsed EPR distance measurements in soluble proteins by site-directed spin labeling (SDSL).
abstract: 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.
authors: Department of Chemistry, University of Florida, Gainesville, Florida.
authors: Ian Mitchelle S
authors: IMS
authors: de Vera
authors: Department of Chemistry, University of Florida, Gainesville, Florida.|Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, Massachusetts.
authors: Mandy E
authors: ME
authors: Blackburn
authors: Department of Chemistry, University of Florida, Gainesville, Florida.|Syngenta Crop Protection, Minnetonka, Minnesota.
authors: Luis
authors: L
authors: Galiano
authors: Department of Chemistry, University of Florida, Gainesville, Florida.
authors: Gail E
authors: GE
authors: Fanucci
mesh_terms: D004578:Electron Spin Resonance Spectroscopy / Q000295:instrumentation / Q000379:methods; D011506:Proteins / Q000737:chemistry*; D013113:Spin Labels
chemical_list: D011506:Proteins; D013113:Spin Labels
keywords: DEER; distance measurements; pulsed EPR; site-directed spin labeling