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Harnessing photoinduced electron transfer to optically determine protein sub-nanoscale atomic distances
Linköping University, Department of Clinical and Experimental Medicine, Divison of Neurobiology. Linköping University, Faculty of Medicine and Health Sciences. Univ Calif Los Angeles, CA 90095 USA.
Amgen Inc, CA 91320 USA.
Univ Calif Los Angeles, CA 90095 USA.
Univ Calif Los Angeles, CA 90095 USA.
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2018 (English)In: Nature Communications, ISSN 2041-1723, E-ISSN 2041-1723, Vol. 9, article id 4738Article in journal (Refereed) Published
Abstract [en]

Proteins possess a complex and dynamic structure, which is influenced by external signals and may change as they perform their biological functions. We present an optical approach, distance-encoding photoinduced electron transfer (DEPET), capable of the simultaneous study of protein structure and function. An alternative to FRET-based methods, DEPET is based on the quenching of small conjugated fluorophores by photoinduced electron transfer: a reaction that requires contact of the excited fluorophore with a suitable electron donor. This property allows DEPET to exhibit exceptional spatial and temporal resolution capabilities in the range pertinent to protein conformational change. We report the first implementation of DEPET on human large-conductance K+ (BK) channels under voltage clamp. We describe conformational rearrangements underpinning BK channel sensitivity to electrical excitation, in conducting channels expressed in living cells. Finally, we validate DEPET in synthetic peptide length standards, to evaluate its accuracy in measuring sub-and near-nanometer intramolecular distances.

Place, publisher, year, edition, pages
NATURE PUBLISHING GROUP , 2018. Vol. 9, article id 4738
National Category
Biophysics
Identifiers
URN: urn:nbn:se:liu:diva-152803DOI: 10.1038/s41467-018-07218-6ISI: 000449628200009PubMedID: 30413716OAI: oai:DiVA.org:liu-152803DiVA, id: diva2:1265297
Note

Funding Agencies|NIH/NINDS [R21NS101734]; NIH/NIGMS Instrument Supplement [R01GM78844]; American Heart Association [14SDG20300018]; Knut and Alice Wallenberg Foundation

Available from: 2018-11-22 Created: 2018-11-22 Last updated: 2019-03-22

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