en
Scientific article
Review
Open access
English

From in vitro towards in situ : structure‐based investigation of ABC exporters by electron paramagnetic resonance spectroscopy

Published inFEBS letters, vol. 594, no. 23, p. 3839-3856
Publication date2020-12
First online date2020-12
Abstract

ATP‐binding cassette (ABC) exporters have been studied now for more than four decades, and recent structural investigation has produced a large number of protein database entries. Yet, important questions about how ABC exporters function at the molecular level remain debated, such as which are the molecular recognition hotspots and the allosteric couplings dynamically regulating the communication between the catalytic cycle and the export of substrates. This conundrum mainly arises from technical limitations confining all research to in vitro analysis of ABC transporters in detergent solutions or embedded in membrane‐mimicking environments. Therefore, a largely unanswered question is how ABC exporters operate in situ , namely in the native membrane context of a metabolically active cell. This review focuses on novel mechanistic insights into type I ABC exporters gained through a unique combination of structure determination, biochemical characterization, generation of conformation‐specific nanobodies/sybodies and double electron–electron resonance.

eng
Keywords
  • ABC transporters
  • DEER
  • EPR
  • MTSL
  • Detergent
  • Inside-out vesicles
  • Liposomes
  • Nanobody
  • Spin label
  • Sybody
Affiliation Not a UNIGE publication
Citation (ISO format)
BORDIGNON, Enrica et al. From <i>in vitro</i> towards <i>in situ</i> : structure‐based investigation of ABC exporters by electron paramagnetic resonance spectroscopy. In: FEBS letters, 2020, vol. 594, n° 23, p. 3839–3856. doi: 10.1002/1873-3468.14004
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Article (Published version)
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ISSN of the journal0014-5793
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