From invitro towards insitu : structure-based investigation of ABC exporters by electron paramagnetic resonance spectroscopy

FEBS Letters

article

Enrica Bordignon¹  Markus A. Seeger²  Laura Galazzo¹  Gianmarco Meier² 

[1] Faculty of Chemistry and Biochemistry, Ruhr University Bochum;Institute of Medical Microbiology, University of Zurich

关键词: ABC transporters;    DEER;    detergent;    EPR;    inside–out vesicles;    liposomes;    MTSL;    nanobody;    spin label;    sybody;   

DOI  :  10.1002/1873-3468.14004

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown