Structural basis for xenobiotic extrusion by eukaryotic MATE transporter

Miyauchi, Hirotake; Moriyama, Shigeharu; Kusakizako, Tsukasa; Kumazaki, Kaoru; Nakane, Takanori; Yamashita, Keitaro; Hirata, Kunio; Dohmae, Naoshi; Nishizawa, Takashi; Ito, Koichi; Miyaji, Takaaki; Moriyama, Yoshinori; Ishitani, Ryuichiro; Nureki, Osamu

doi:10.1038/s41467-017-01541-0

Cited by 76 publications

(76 citation statements)

References 42 publications

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“…The outward-facing cavity was substantially shallower and narrower than the inward-facing cavity, and thus cannot accommodate the head group of Lipid II. This suggests that our outward facing structure represents a state in which the head group of Lipid II is released into the periplasm, consistent with the idea that cavity shrinkage could be a mechanism to displace substrate into the periplasm 32,34 (Fig. 4a).…”

Section: Outward-facing Conformation Of Murjtasupporting

confidence: 84%

“…Furthermore, the questions of whether MurJ is an ion-coupled transporter and which ion(s) are involved remain unresolved. While some drug exporters in the MOP superfamily have been characterized to be Na + or H + coupled 12,[30][31][32] , our high-resolution inward-facing MurJ structure did not reveal a cation that could be important for transport 25 . It has been shown that MurJ activity is dependent on membrane potential and not on a H + gradient 33 , but the involvement of Na + has not been ruled out.…”

Section: Main Textmentioning

confidence: 57%

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Visualizing Conformation Transitions of the Lipid Flippase MurJ

Kuk

Hao

Guan

et al. 2019

Preprint

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The biosynthesis of many polysaccharides, including bacterial peptidoglycan and eukaryotic N-linked glycans, requires transport of lipid-linked oligosaccharide (LLO) precursors across the membrane by specialized flippases. MurJ is the flippase for the lipid-linked peptidoglycan precursor Lipid II, a key player in bacterial cell wall synthesis, and a target of recently discovered antibacterials. However, the flipping mechanism of LLOs including Lipid II remains poorly understood due to a dearth of structural information. Here we report crystal structures of MurJ captured in inward-closed, inward-open, inward-occluded and outward-facing conformations. Together with cysteine accessibility, mass spectrometry, and complementation studies, we elucidate the conformational transitions in MurJ that mediate lipid flipping, identified the key ion for function, and provide a framework for the development of inhibitors.

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Section: Outward-facing Conformation Of Murjtasupporting

confidence: 84%

Section: Main Textmentioning

confidence: 57%

Visualizing Conformation Transitions of the Lipid Flippase MurJ

Kuk

Hao

Guan

et al. 2019

Preprint

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“…the NorM, DinF, and eukaryotic subfamilies (Omote et al, 2006). Over the last decade, structural and biochemical analyses of MATE transporters have been intensively pursued (Miyauchi et al, 2017;Tanaka et al, 2013Tanaka et al, , 2017He et al, 2010;Lu et al, 2013aLu et al, , 2013bMousa et al, 2016;Radchenko et al, 2015). To date, the crystal structures of five prokaryotic MATE transporters have been reported: Vibrio cholerae NorM subfamily (NorM-VC) (He et al, 2010) simultaneously coupled to the Na + and H + gradients (Jin et al, 2014), two Na + -driven MATE transporters (Neisseria gonorrhoeae NorM [NorM-NG] [Lu et al, 2013a] and Escherichia coli DinF subfamilies [ClbM] [Mousa et al, 2016]), and two H + -driven MATE transporters (Pyrococcus furiosus DinF [PfMATE] and Bacillus halodurans DinF subfamilies [DinF-BH] [Lu et al, 2013b;Radchenko et al, 2015]).…”

Section: Introductionmentioning

confidence: 99%

Structural Basis of H+-Dependent Conformational Change in a Bacterial MATE Transporter

et al. 2019

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Graphical Abstract Highlights d Crystal structures of VcmN, an H + -coupled MATE transporter from Vibrio cholerae d Distinct conformations associated with different pHs and the D35N mutation in TM1 d Coupling of rearrangement of hydrogen bond network around Asp35 to the TM1 bending d Common step in the transport cycle shared among prokaryotic H + -driven MATE proteins SUMMARY Multidrug and toxic compound extrusion (MATE) transporters efflux toxic compounds using a Na + or H + gradient across the membrane. Although the structures of MATE transporters have been reported, the cation-coupled substrate transport mechanism remains controversial. Here we report crystal structures of VcmN, a Vibrio cholerae MATE transporter driven by the H + gradient. High-resolution structures in two distinct conformations associated with different pHs revealed that the rearrangement of the hydrogen-bonding network around the conserved Asp35 induces the bending of transmembrane helix 1, as in the case of the H + -coupled Pyrococcus furiosus MATE transporter. We also determined the crystal structure of the D35N mutant, which captured a unique conformation of TM1 facilitated by an altered hydrogen-bonding network. Based on the present results, we propose a common step in the transport cycle shared among prokaryotic H + -coupled MATE transporters.

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“…Based on their amino acid sequence similarity, the MATE family members are classified into the NorM, the DNA damage-inducible protein F (DinF), and the eukaryotic subfamilies (2). In recent years, the crystal structures of representative members of all three subfamilies have been published (7)(8)(9)(10)(11)(12)(13)(14)(15). Since all of them represent only an outward-facing state, a detailed understanding of the complete transport cycle has remained elusive.…”

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confidence: 99%

Inward-facing conformation of a multidrug resistance MATE family transporter

Zakrzewska

Mehdipour

Malviya

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

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Multidrug and toxic compound extrusion (MATE) transporters mediate excretion of xenobiotics and toxic metabolites, thereby conferring multidrug resistance in bacterial pathogens and cancer cells. Structural information on the alternate conformational states and knowledge of the detailed mechanism of MATE transport are of great importance for drug development. However, the structures of MATE transporters are only known in V-shaped outward-facing conformations. Here, we present the crystal structure of a MATE transporter from Pyrococcus furiosus (PfMATE) in the long-soughtafter inward-facing state, which was obtained after crystallization in the presence of native lipids. Transition from the outward-facing state to the inward-facing state involves rigid body movements of transmembrane helices (TMs) 2-6 and 8-12 to form an inverted V, facilitated by a loose binding of TM1 and TM7 to their respective bundles and their conformational flexibility. The inward-facing structure of PfMATE in combination with the outward-facing one supports an alternating access mechanism for the MATE family transporters. multidrug resistance | membrane protein structure | MATE transporter | inward-facing conformation | lipids

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Structural basis for xenobiotic extrusion by eukaryotic MATE transporter

Cited by 76 publications

References 42 publications

Visualizing Conformation Transitions of the Lipid Flippase MurJ

Visualizing Conformation Transitions of the Lipid Flippase MurJ

Structural Basis of H+-Dependent Conformational Change in a Bacterial MATE Transporter

Inward-facing conformation of a multidrug resistance MATE family transporter

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