Single-molecule probing of the conformational homogeneity of the ABC transporter BtuCD

Min Yang; Nurit Livnat Levanon; Burçin Acar; Burcu Aykac Fas; Gal Masrati; Jessica Rose; Nir Ben-Tal; Turkan Haliloglu; Yongfang Zhao; Oded Lewinson

doi:10.1038/s41589-018-0088-2

Single-molecule probing of the conformational homogeneity of the ABC transporter BtuCD

Min Yang, Nurit Livnat Levanon, Burçin Acar, Burcu Aykac Fas, Gal Masrati, Jessica Rose, Nir Ben-Tal, Turkan Haliloglu, Yongfang Zhao, Oded Lewinson

Research output: Contribution to journal › Article › peer-review

40 Scopus citations

Abstract

ATP-binding cassette (ABC) transporters use the energy of ATP hydrolysis to move molecules through cellular membranes. They are directly linked to human diseases, cancer multidrug resistance, and bacterial virulence. Very little is known of the conformational dynamics of ABC transporters, especially at the single-molecule level. Here, we combine single-molecule spectroscopy and a novel molecular simulation approach to investigate the conformational dynamics of the ABC transporter BtuCD. We observe a single dominant population of molecules in each step of the transport cycle and tight coupling between conformational transitions and ligand binding. We uncover transient conformational changes that allow substrate to enter the transporter. This is followed by a 'squeezing' motion propagating from the extracellular to the intracellular side of the translocation cavity. This coordinated sequence of events provides a mechanism for the unidirectional transport of vitamin B ₁₂ by BtuCD.

Original language	English
Pages (from-to)	715-722
Number of pages	8
Journal	Nature Chemical Biology
Volume	14
Issue number	7
DOIs	https://doi.org/10.1038/s41589-018-0088-2
State	Published - 1 Jul 2018
Externally published	Yes

Bibliographical note

Publisher Copyright:
© 2018 The Author(s).

Funding

This work is in honor and memory of Yongfang Zhao.The authors thank B. Poolman and A. Meller for fruitful discussions. The plasmid for Cys-less BtuCD was a kind gift from K. Locher (ETH Zurich). This work was supported by grants from NATO Science for Peace and Security Program (SPS Project G4622, O.L., N.L.L., J.R., T.H., B.A., B.A.F., N.B.T., and G.M.), the Israeli Academy of Sciences (O.L., N.L.L.), TUBITAK (The Scientific and Technological Research Council of Turkey) under the grant no 115M418 (T.H., B.A., B.A.F.), the Rappaport Family Institute for biomedical research (O.L., N.L.L., J.R.), the Ministry of Science and Technology (China) “973” Project grant 2014CB910400 (M.Y., J.Z.), the National Natural Science Foundation of China (31522016) (M.Y. and Y.Z.), and the Merieux research foundation (O.L., N.L.L.).

Funders	Funder number
Israeli Academy of Sciences
Merieux research foundation
Ministry of Science and Technology (China) “973” Project	2014CB910400
NATO Science for Peace and Security Program
Rappaport Family Institute
TUBITAK
National Natural Science Foundation of China	31522016
Saudi Pharmaceutical Society	G4622
Türkiye Bilimsel ve Teknolojik Araştirma Kurumu	115M418

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abstract = " ATP-binding cassette (ABC) transporters use the energy of ATP hydrolysis to move molecules through cellular membranes. They are directly linked to human diseases, cancer multidrug resistance, and bacterial virulence. Very little is known of the conformational dynamics of ABC transporters, especially at the single-molecule level. Here, we combine single-molecule spectroscopy and a novel molecular simulation approach to investigate the conformational dynamics of the ABC transporter BtuCD. We observe a single dominant population of molecules in each step of the transport cycle and tight coupling between conformational transitions and ligand binding. We uncover transient conformational changes that allow substrate to enter the transporter. This is followed by a 'squeezing' motion propagating from the extracellular to the intracellular side of the translocation cavity. This coordinated sequence of events provides a mechanism for the unidirectional transport of vitamin B 12 by BtuCD.",

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AU - Masrati, Gal

AU - Rose, Jessica

AU - Ben-Tal, Nir

AU - Haliloglu, Turkan

AU - Zhao, Yongfang

AU - Lewinson, Oded

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AB - ATP-binding cassette (ABC) transporters use the energy of ATP hydrolysis to move molecules through cellular membranes. They are directly linked to human diseases, cancer multidrug resistance, and bacterial virulence. Very little is known of the conformational dynamics of ABC transporters, especially at the single-molecule level. Here, we combine single-molecule spectroscopy and a novel molecular simulation approach to investigate the conformational dynamics of the ABC transporter BtuCD. We observe a single dominant population of molecules in each step of the transport cycle and tight coupling between conformational transitions and ligand binding. We uncover transient conformational changes that allow substrate to enter the transporter. This is followed by a 'squeezing' motion propagating from the extracellular to the intracellular side of the translocation cavity. This coordinated sequence of events provides a mechanism for the unidirectional transport of vitamin B 12 by BtuCD.

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Single-molecule probing of the conformational homogeneity of the ABC transporter BtuCD

Abstract

Bibliographical note

Funding

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