Exploring Structure, Dynamics, and Topology of Nitroxide Spin-Labeled Proteins Using Continuous-Wave Electron Paramagnetic Resonance Spectroscopy

Christian Altenbach, Carlos J. López, Kálmán Hideg, Wayne L. Hubbell

Research output: Chapter in Book/Report/Conference proceedingChapter

35 Citations (Scopus)

Abstract

Structural and dynamical characterization of proteins is of central importance in understanding the mechanisms underlying their biological functions. Site-directed spin labeling (SDSL) combined with continuous-wave electron paramagnetic resonance (CW EPR) spectroscopy has shown the capability of providing this information with site-specific resolution under physiological conditions for proteins of any degree of complexity, including those associated with membranes. This chapter introduces methods commonly employed for SDSL and describes selected CW EPR-based methods that can be applied to (1) map secondary and tertiary protein structure, (2) determine membrane protein topology, (3) measure protein backbone flexibility, and (4) reveal the existence of conformational exchange at equilibrium.

Original languageEnglish
Title of host publicationMethods in Enzymology
PublisherAcademic Press Inc.
Pages59-100
Number of pages42
DOIs
Publication statusPublished - Jan 1 2015

Publication series

NameMethods in Enzymology
Volume564
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

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Keywords

  • CW EPR
  • Distance measurements
  • EPR lineshape analysis
  • Nitroxide scanning
  • Protein dynamics
  • Protein structure
  • Site-directed spin labeling
  • Solvent accessibility
  • Spin interaction

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Altenbach, C., López, C. J., Hideg, K., & Hubbell, W. L. (2015). Exploring Structure, Dynamics, and Topology of Nitroxide Spin-Labeled Proteins Using Continuous-Wave Electron Paramagnetic Resonance Spectroscopy. In Methods in Enzymology (pp. 59-100). (Methods in Enzymology; Vol. 564). Academic Press Inc.. https://doi.org/10.1016/bs.mie.2015.08.006