Crystal structure of the MOP flippase MurJ in an inward-facing conformation

Kuk, Alvin C. Y.; Mashalidis, Ellene H.; Lee, Seok‐Yong

doi:10.1038/nsmb.3346

“…Moreover, it is related to transporters implicated in translocating other lipid-linked polysaccharide precursors such as those for O-antigen synthesis 9 . Finally, chemical probing and a recently published crystal structure indicate that MurJ has a solvent-exposed cavity consistent with it functioning as a transporter 10,11. …”

mentioning

confidence: 68%

“…The Lys M -resistant mutants were rendered on the crystal structure of MurJ TA (PDB 5T77) 11 from Thermsipho africanus or the outward-facing model of MurJ TA 11 using the UCSF Chimera package 26 .…”

Section: Methodsmentioning

confidence: 99%

“…The model presented here was the model with the highest level of confidence scores (C-score −0.17; TM-score= 0.69±0.12). The top-ranked structural analogue to the model generated by I-TASSER is the recently published structure of the MurJ homologue from Thermosipho africanus (PDB 5T77) 11 , and comparison of these two structures resulted in a TM-score of 0.894, where 1 indicates a perfect match. Figures were prepared from PBD files downloaded from the I-TASSER server using the PyMOL Molecular Graphics System, Version 1.5.0.4 (Schrödinger, LLC).…”

Section: Methodsmentioning

confidence: 99%

“…The amino-acid changes in E. coli MurJ (MurJ EC ) resulting in Lys M -resistance were mapped onto the structure of MurJ from Thermosipho africanus (MurJ TA ) (PDB 5T77) 11 . a , The cytoplasmic-open conformation.…”

Section: Figmentioning

confidence: 99%

“…a , The cytoplasmic-open conformation. b , A model of the periplasmic-open conformation 11 . The TMDs that line the central hydrophilic cavity are coloured: TMD2 (light blue) and TMD7 (magenta).…”

Section: Figmentioning

confidence: 99%

See 3 more Smart Citations

A viral protein antibiotic inhibits lipid II flippase activity

Chamakura

¹

,

Sham

²

,

Davis

³

et al. 2017

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For bacteriophage infections, the cell walls of bacteria, consisting of a single highly polymeric molecule of peptidoglycan (PG), pose a major problem for the release of progeny virions. Phage lysis proteins that overcome this barrier can point the way to new antibacterial strategies1, especially small lytic single-stranded DNA (the microviruses) and RNA phages (the leviviruses) that effect host lysis using a single non-enzymatic protein2. Previously, the A2 protein of levivirus Qβ and the E protein of the microvirus φ X174 were shown to be ‘protein antibiotics’ that inhibit the MurA and MraY steps of the PG synthesis pathway2–4. Here, we investigated the mechanism of action of an unrelated lysis protein, LysM, of the Escherichia coli levivirus M5. We show that LysM inhibits the translocation of the final lipid-linked PG precursor called lipid II across the cytoplasmic membrane by interfering with the activity of MurJ. The finding that LysM inhibits a distinct step in the PG synthesis pathway from the A2 and E proteins indicates that small phages, particularly the single-stranded RNA (ssRNA) leviviruses, have a previously unappreciated capacity for evolving novel inhibitors of PG biogenesis despite their limited coding potential.

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A Mass‐Spectrometry‐Based Approach to Distinguish Annular and Specific Lipid Binding to Membrane Proteins

Bolla

¹

,

Corey

²

,

Sahin

³

et al. 2020

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Membrane proteins engage in a variety of contacts with their surrounding lipids, but distinguishing between specifically bound lipids, and non‐specific, annular interactions is a challenging problem. Applying native mass spectrometry to three membrane protein complexes with different lipid‐binding properties, we explore the ability of detergents to compete with lipids bound in different environments. We show that lipids in annular positions on the presenilin homologue protease are subject to constant exchange with detergent. By contrast, detergent‐resistant lipids bound at the dimer interface in the leucine transporter show decreased koff rates in molecular dynamics simulations. Turning to the lipid flippase MurJ, we find that addition of the natural substrate lipid‐II results in the formation of a 1:1 protein–lipid complex, where the lipid cannot be displaced by detergent from the highly protected active site. In summary, we distinguish annular from non‐annular lipids based on their exchange rates in solution.

show abstract

A Mass‐Spectrometry‐Based Approach to Distinguish Annular and Specific Lipid Binding to Membrane Proteins

Bolla

¹

,

Corey

²

,

Sahin

³

et al. 2020

View full text Add to dashboard Cite

Membrane proteins engage in a variety of contacts with their surrounding lipids, but distinguishing between specifically bound lipids, and non‐specific, annular interactions is a challenging problem. Applying native mass spectrometry to three membrane protein complexes with different lipid‐binding properties, we explore the ability of detergents to compete with lipids bound in different environments. We show that lipids in annular positions on the presenilin homologue protease are subject to constant exchange with detergent. By contrast, detergent‐resistant lipids bound at the dimer interface in the leucine transporter show decreased koff rates in molecular dynamics simulations. Turning to the lipid flippase MurJ, we find that addition of the natural substrate lipid‐II results in the formation of a 1:1 protein–lipid complex, where the lipid cannot be displaced by detergent from the highly protected active site. In summary, we distinguish annular from non‐annular lipids based on their exchange rates in solution.

show abstract

Crystal structure of the MOP flippase MurJ in an inward-facing conformation

Cited by 76 publications

References 33 publications

A viral protein antibiotic inhibits lipid II flippase activity

A viral protein antibiotic inhibits lipid II flippase activity

A Mass‐Spectrometry‐Based Approach to Distinguish Annular and Specific Lipid Binding to Membrane Proteins

A Mass‐Spectrometry‐Based Approach to Distinguish Annular and Specific Lipid Binding to Membrane Proteins

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