Unit

UNIT 17.17 Pulsed EPR Distance Measurements in Soluble Proteins by Site-Directed Spin Labeling (SDSL)

  1. Ian Mitchelle S. de Vera1,
  2. Mandy E. Blackburn1,2,
  3. Luis Galiano1,3,
  4. Gail E. Fanucci1

Published Online: 5 NOV 2013

DOI: 10.1002/0471140864.ps1717s74

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

de Vera, I. M. S., Blackburn, M. E., Galiano, L. and Fanucci, G. E. 2013. Pulsed EPR Distance Measurements in Soluble Proteins by Site-Directed Spin Labeling (SDSL). Current Protocols in Protein Science. 74:17.17:17.17.1–17.17.29.

Author Information

  1. 1

    Department of Chemistry, University of Florida, Gainesville, Florida

  2. 2

    Department of Biochemistry and Molecular Biology, University of Massachusetts Amherst, Amherst, Massachusetts

  3. 3

    Syngenta Crop Protection, Minnetonka, Minnesota

Publication History

  1. Published Online: 5 NOV 2013

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. Curr. Protoc. Protein Sci. 74:17.17.1-17.17.29. © 2013 by John Wiley & Sons, Inc.

Keywords:

  • pulsed EPR;
  • DEER;
  • distance measurements;
  • site-directed spin labeling