Format

Send to

Choose Destination
Biophys J. 2013 Dec 17;105(12):2724-32. doi: 10.1016/j.bpj.2013.11.010.

Tethered spectroscopic probes estimate dynamic distances with subnanometer resolution in voltage-dependent potassium channels.

Author information

1
Department of Neuroscience, University of Wisconsin, Madison, Wisconsin.
2
School of Pharmacy, Division of Pharmaceutical Sciences, University of Wisconsin, Madison, Wisconsin.
3
Department of Chemistry and Theoretical Chemistry Institute, University of Wisconsin, Madison, Wisconsin.
4
Department of Neuroscience, University of Wisconsin, Madison, Wisconsin. Electronic address: chanda@wisc.edu.

Abstract

Measurements of inter- and intramolecular distances are important for monitoring structural changes and understanding protein interaction networks. Fluorescence resonance energy transfer and functionalized chemical spacers are the two predominantly used strategies to map short-range distances in living cells. Here, we describe the development of a hybrid approach that combines the key advantages of spectroscopic and chemical methods to estimate dynamic distance information from labeled proteins. Bifunctional spectroscopic probes were designed to make use of adaptable-anchor and length-varied spacers to estimate molecular distances by exploiting short-range collisional electron transfer. The spacers were calibrated using labeled polyproline peptides of defined lengths and validated by molecular simulations. This approach was extended to estimate distance restraints that enable us to evaluate the resting-state model of the Shaker potassium channel.

Comment in

PMID:
24359744
PMCID:
PMC3882458
DOI:
10.1016/j.bpj.2013.11.010
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center