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Updated: May 17, 2026
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Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR
Published on: December 16, 2013
Nano-positioning system for structural analysis of functional homomeric proteins in multiple conformations
H Clark Hyde1, Walter Sandtner, Ernesto Vargas
1Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA. fbezanilla@uchicago.edu
Structure (London, England : 1993)
|October 16, 2012
Summary
This study introduces a new method to map protein structures using inverse trilateration and resonance energy transfer (RET). The technique accurately determines protein conformations, including those not previously visualized.
Area of Science:
- Biophysics
- Structural Biology
- Biochemistry
Background:
- Proteins undergo conformational changes crucial for their function.
- Resonance energy transfer (RET) measures distances between labeled sites to infer protein structure.
- Interpreting RET data in complex three-dimensional rearrangements, especially in homomeric proteins, is challenging.
Purpose of the Study:
- To develop a novel positioning method for mapping target sites within homomeric proteins across multiple conformational states.
- To accurately determine the three-dimensional positions and confidence regions of labeled sites using inverse trilateration/triangulation.
- To simultaneously record protein function alongside structural determination.
Main Methods:
- Developed a positioning method employing inverse trilateration/triangulation.
- Utilized lanthanide RET with donors on target sites (one per subunit) and a single acceptor on a static site.
- Accounted for probe diffusion to enhance positional accuracy.
- Applied the method to a functional voltage-gated potassium channel.
Main Results:
- Successfully mapped target sites within a homomeric protein in defined states.
- Accurately determined the 3D position and confidence region of lanthanide LRET donors.
- Verified the crystal structure's relaxed conformation of a voltage-gated potassium channel.
- Reported on the resting and active conformations of the channel, for which crystal structures were unavailable.
Conclusions:
- The developed inverse trilateration/triangulation method accurately maps protein structures in multiple conformations.
- This technique is particularly valuable for studying homomeric proteins and their dynamic rearrangements.
- The study provides new structural insights into voltage-gated potassium channel conformations.
