Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Identifying conformational changes with site-directed spin labeling.

W L Hubbell1, D S Cafiso, C Altenbach

  • 1Jules Stein Eye Institute and Department of Chemistry and Biochemistry, University of California, Los Angeles, California 90095, USA. hubbellw@jsei.ucla.edu

Nature Structural Biology
|August 31, 2000
PubMed
Summary

Site-directed spin labeling and electron paramagnetic resonance (EPR) spectroscopy reveal protein structural changes. This method tracks protein dynamics and rearrangements, offering insights into molecular function.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Paramagnetic hydrophobic ions as probes for electrically active conformational transitions in ion channels.

Biophysical journal·2009
Same author

Structure and dynamics of annexin 12 bound to a planar lipid bilayer.

Physical review letters·2003
Same author

Estimation of inter-residue distances in spin labeled proteins at physiological temperatures: experimental strategies and practical limitations.

Biochemistry·2001
Same author

Structure and function in rhodopsin: mapping light-dependent changes in distance between residue 65 in helix TM1 and residues in the sequence 306-319 at the cytoplasmic end of helix TM7 and in helix H8.

Biochemistry·2001
Same author

Structure and function in rhodopsin: mapping light-dependent changes in distance between residue 316 in helix 8 and residues in the sequence 60-75, covering the cytoplasmic end of helices TM1 and TM2 and their connection loop CL1.

Biochemistry·2001
Same author

Transport-defective mutations alter the conformation of the energy-coupling motif of an outer membrane transporter.

Biochemistry·2001

Area of Science:

  • Biochemistry
  • Structural Biology
  • Spectroscopy

Background:

  • Protein structure and dynamics are crucial for biological function.
  • Electron paramagnetic resonance (EPR) spectroscopy is a valuable technique for studying molecular systems.
  • Site-directed spin labeling (SDSL) allows for specific probing of protein environments.

Purpose of the Study:

  • To review strategies for interpreting SDSL-EPR data in terms of protein structural changes.
  • To highlight the capabilities of SDSL-EPR for studying protein dynamics and function.

Main Methods:

  • Site-directed spin labeling of proteins.
  • Electron paramagnetic resonance (EPR) spectroscopy.
  • Analysis of spin label mobility, solvent accessibility, and inter-label distances.

Related Experiment Videos

Main Results:

  • SDSL-EPR can detect rigid body motions of secondary structure elements (alpha-helices, beta-strands).
  • The technique reveals relative domain movements and changes in secondary structure.
  • Structural changes can be monitored on the millisecond timescale, enabling functional studies.

Conclusions:

  • SDSL-EPR is a versatile and powerful method for investigating protein structure and dynamics.
  • It is applicable to proteins of all sizes and requires only small amounts of protein.
  • This approach facilitates the study of a wide range of biological systems.