Structure of a Type-1 Secretion System ABC Transporter

Morgan, Jacob L. W.; Acheson, Justin; Zimmer, Jochen

doi:10.1016/j.str.2017.01.010

Cited by 71 publications

(56 citation statements)

References 45 publications

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“…The TMD is occluded to either side of the membrane in the presence or absence of nucleotides that was also supported by Pulsed Electron-Electron Double Resonance (PELDOR) measurements in bicelles (Bountra et al, 2017). Occluded conformations have also been reported for other ABC transporters (Lin et al, 2015;Perez et al, 2015;Morgan et al, 2017). We have biochemically shown that McjD adopts a transient outward-open conformation that it is probably not long-lived or well populated to be trapped by PELDOR (Bountra et al, 2017).…”

Section: Introductionsupporting

confidence: 66%

Conformational dynamics of the ABC transporter McjD seen by single‐molecule FRET

et al. 2018

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ABC transporters utilize ATP for export processes to provide cellular resistance against toxins, antibiotics, and harmful metabolites in eukaryotes and prokaryotes. Based on static structure snapshots, it is believed that they use an alternating access mechanism, which couples conformational changes to ATP binding (outward‐open conformation) and hydrolysis (inward‐open) for unidirectional transport driven by ATP. Here, we analyzed the conformational states and dynamics of the antibacterial peptide exporter McjD from Escherichia coli using single‐molecule Förster resonance energy transfer (smFRET). For the first time, we established smFRET for an ABC exporter in a native‐like lipid environment and directly monitor conformational dynamics in both the transmembrane‐ (TMD) and nucleotide‐binding domains (NBD). With this, we unravel the ligand dependences that drive conformational changes in both domains. Furthermore, we observe intrinsic conformational dynamics in the absence of ATP and ligand in the NBDs. ATP binding and hydrolysis on the other hand can be observed via NBD conformational dynamics. We believe that the progress made here in combination with future studies will facilitate full understanding of ABC transport cycles.

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Section: Introductionsupporting

confidence: 66%

Conformational dynamics of the ABC transporter McjD seen by single‐molecule FRET

et al. 2018

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“…9, C and D, yellow circles). In addition, on the kinked helix on the substrate entry window, there is a positively-charged residue that sticks out toward the pore of the window and blocks the window in ADP-bound state of TliD (34). The ConSurf results also verified that this residue was charge-conserved, as all of the 50 homologs had either arginine or lysine at this residue (Fig.…”

Section: Protein Isoelectric Point and Abc Transporter Secretionmentioning

confidence: 72%

A lower isoelectric point increases signal sequence–mediated secretion of recombinant proteins through a bacterial ABC transporter

Byun

Park

Kim

et al. 2017

Journal of Biological Chemistry

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Efficient protein production for industrial and academic purposes often involves engineering microorganisms to produce and secrete target proteins into the culture. has a TliDEF ATP-binding cassette transporter, a type I secretion system, which recognizes C-terminal LARD3 signal sequence of thermostable lipase TliA. Many proteins are secreted by TliDEF when recombined with LARD3, but there are still others that cannot be secreted by TliDEF even when LARD3 is attached. However, the factors that determine whether or not a recombinant protein can be secreted through TliDEF are still unknown. Here, we recombined LARD3 with several proteins and examined their secretion through TliDEF. We found that the proteins secreted via LARD3 are highly negatively charged with highly-acidic isoelectric points (pI) lower than 5.5. Attaching oligo-aspartate to lower the pI of negatively-charged recombinant proteins improved their secretion, and attaching oligo-arginine to negatively-charged proteins blocked their secretion by LARD3. In addition, negatively supercharged green fluorescent protein (GFP) showed improved secretion, whereas positively supercharged GFP did not secrete. These results disclosed that proteins' acidic pI and net negative charge are major factors that determine their secretion through TliDEF. Homology modeling for TliDEF revealed that TliD dimer forms evolutionarily-conserved positively-charged clusters in its pore and substrate entrance site, which also partially explains the pI dependence of the TliDEF-dependent secretions. In conclusion, lowering the isoelectric point improved LARD3-mediated protein secretion, both widening the range of protein targets for efficient production via secretion and signifying an important aspect of ABC transporter-mediated secretions.

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“…In this state, PrtD binds the other components PrtE and PrtF [77,[110][111][112]. The full assembly is suggested to reactivate PrtD's ATPase activity and therewith to enable the substrate transport [77,113,114]. A potentially related stimulation by the MFP has previously been demonstrated for the interaction of MacA with MacB [61].…”

Section: Macb and Type I Secretion System Abc Transportersmentioning

confidence: 99%

“…The recently solved structures of the inner membrane ABC transporter component of a type I secretion system (Aquifex aeolicus AaPrtD, part of AaPrtDEF assembly) [77] and that of the ABC transporter MacB from the tripartite multi-drug efflux assembly MacAB-TolC [33,[37][38][39]] invite a structural comparison of these two systems. Both structures are depicted in Fig.…”

Section: Macb and Type I Secretion System Abc Transportersmentioning

confidence: 99%

“…In the case of PrtD, a basal ATPase activity may be repressed by substrate binding, which could prevent the closure of nucleotide-binding domains. In this state, PrtD binds the other components PrtE and PrtF [77,[110][111][112]. The full assembly is suggested to reactivate PrtD's ATPase activity and therewith to enable the substrate transport [77,113,114].…”

Section: Macb and Type I Secretion System Abc Transportersmentioning

confidence: 99%

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Structure and mechanism of bacterial tripartite efflux pumps

Neuberger

Luisi

2018

Research in Microbiology

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Efflux pumps are membrane proteins which contribute to multi-drug resistance. In Gram-negative bacteria, some of these pumps form complex tripartite assemblies in association with an outer membrane channel and a periplasmic membrane fusion protein. These tripartite machineries span both membranes and the periplasmic space, and they extrude from the bacterium chemically diverse toxic substrates. In this chapter, we summarise current understanding of the structural architecture, functionality, and regulation of tripartite multi-drug efflux assemblies.

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Structure of a Type-1 Secretion System ABC Transporter

Cited by 71 publications

References 45 publications

Conformational dynamics of the ABC transporter McjD seen by single‐molecule FRET

Conformational dynamics of the ABC transporter McjD seen by single‐molecule FRET

A lower isoelectric point increases signal sequence–mediated secretion of recombinant proteins through a bacterial ABC transporter

Structure and mechanism of bacterial tripartite efflux pumps

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