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Probing Protein Secondary Structure Using EPR: Investigating a Dynamic Region of Visual Arrestin

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Abstract

One key application of site-directed spin labeling electron paramagnetic resonance (EPR) spectroscopy is the determination of sequence-specific secondary structure in proteins. Regular secondary structure leads to a periodic variation in both side chain motion and solvent accessibility, two properties easily monitored by EPR techniques. Specifically, saturation recovery (SR) EPR spectroscopy has proven to be useful for making accessibility measurements for multiple protein structure populations by determining individual accessibilities and is, therefore, well suited to study the structure of proteins exhibiting multiple conformations in equilibrium. Here we employ both continuous wave and SR EPR spectroscopy in combination to examine the secondary structure of a short sequence showing conformational heterogeneity in visual rod arrestin. The EPR data presented here clearly distinguish between the unstructured loop and the helical structure formed in the crystallographic tetramer of visual arrestin and show that this region is unstructured in solution.

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Authors and Affiliations

  1. Department of Biophysics, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI, 53226, USA

    Derek J. Francis & Candice S. Klug

  2. Department of Chemistry and Biochemistry, Jules Stein Eye Institute, University of California, Los Angeles, CA, 90095, USA

    Wayne L. Hubbell

Authors
  1. Derek J. Francis
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  2. Wayne L. Hubbell
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  3. Candice S. Klug
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Correspondence to Candice S. Klug.

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Francis, D.J., Hubbell, W.L. & Klug, C.S. Probing Protein Secondary Structure Using EPR: Investigating a Dynamic Region of Visual Arrestin. Appl Magn Reson 43, 405–419 (2012). https://doi.org/10.1007/s00723-012-0369-y

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  • DOI: https://doi.org/10.1007/s00723-012-0369-y

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