Dynamic Interaction of the Escherichia coli Cell Division ZipA and FtsZ Proteins Evidenced in Nanodiscs

Hernández-Rocamora, Víctor M.; Reija, Belén; Garcı́a, Concepción; Natale, Paolo; Alfonso, Carlos; Minton, Allen P.; Zorrilla, Silvia; Rivas, Germán; Vicente, Miguel

doi:10.1074/jbc.m112.388959

Cited by 45 publications

(47 citation statements)

References 24 publications

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“…This peptide has been previously shown to inhibit the binding of FtsZ polymers to ZipA incorporated in nanodiscs (27). The presence of 24 M CTZMUT prevented the collapse of vesicles containing 5 M ZipA and 12 M FtsZ, without altering FtsZ polymerization inside the vesicles once GTP and magnesium were added (Fig.…”

Section: Vesicle Shrinkage Is Prevented By An Inhibitor Of the Bindinmentioning

confidence: 99%

“…The peptide CTZ-MUT, with sequence (KRNDWT-NIMAFLKKQAD) (26), was synthesized as described in Ref. 27. The pore-forming protein ␣-hemolysin from Staphylococcus aureus was obtained from Sigma-Aldrich (H9395).…”

Section: Methodsmentioning

confidence: 99%

“…These protein concentrations are close to the estimated physiological concentrations of FtsZ (2-10 M (34)) and ZipA (0.5-1.0 M (34, 39)) in E. coli. Moreover, they are in the range of the FtsZ-ZipA binding affinities obtained for either the soluble ZipA variant (25) or the fulllength version of the protein incorporated in nanodiscs (27). In all cases addition of external GTP and magnesium was used to allow polymerization of FtsZ inside the vesicles.…”

Section: Guvs With Zipa Inserted In the Membrane Shrink Uponmentioning

confidence: 99%

“…The incorporation of the full-length ZipA protein into the vesicles, although technically feasible (38), was not considered because it requires the presence of detergents during the vesicle production that alter the properties of FtsZ. The sZipA variant has the same FtsZ binding properties as the full-length protein incorporated in phospholipid bilayer nanodiscs (25,27).…”

Section: Guvs With Zipa Inserted In the Membrane Shrink Uponmentioning

confidence: 99%

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Bacterial Division Proteins FtsZ and ZipA Induce Vesicle Shrinkage and Cell Membrane Invagination

Cabré

Sánchez-Gorostiaga

Carrara

et al. 2013

Journal of Biological Chemistry

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Background: Before constriction ZipA anchors FtsZ to the E. coli inner membrane as part of the cell division proto-ring. Results: Dynamic FtsZ polymers shrink ZipA-containing vesicles whereas excess of ZipA invaginates the E. coli membrane destroying the permeability barrier. Conclusion: Constriction forces can be evidenced both in bacteria and in vesicles. Significance: Defined bacterial elements reproduce division functions when assembled in vitro.

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Section: Vesicle Shrinkage Is Prevented By An Inhibitor Of the Bindinmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Guvs With Zipa Inserted In the Membrane Shrink Uponmentioning

confidence: 99%

Section: Guvs With Zipa Inserted In the Membrane Shrink Uponmentioning

confidence: 99%

See 2 more Smart Citations

Bacterial Division Proteins FtsZ and ZipA Induce Vesicle Shrinkage and Cell Membrane Invagination

Cabré

Sánchez-Gorostiaga

Carrara

et al. 2013

Journal of Biological Chemistry

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“…A variety of synthetic lipids have been used for nanodisc preparation, including variants of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylglycerol (PG), as well as Escherichia coli lipid extracts (25,(27)(28)(29). Diverse membrane proteins have been incorporated into nanodiscs and studies on these systems have revealed fascinating nuances of protein structure, protein oligomeric state, protein-protein interactions, substrate binding, and enzyme activity (30)(31)(32)(33)(34)(35)(36). In addition, particularly noteworthy for the current study, the effects of lipid composition on enzymes have been probed in a highly reproducible manner relative to other membrane mimetic systems (37,38).…”

mentioning

confidence: 99%

Lipid bilayer nanodisc platform for investigating polyprenol-dependent enzyme interactions and activities

Hartley

Schneggenburger

Imperiali

2013

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

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Membrane-bound polyprenol-dependent pathways are important for the assembly of essential glycoconjugates in all domains of life. However, despite their prevalence, the functional significance of the extended linear polyprenyl groups in the interactions of the glycan substrates, the biosynthetic enzymes that act upon them, and the membrane bilayer in which they are embedded remains a mystery. These interactions are investigated simultaneously and uniquely through application of the nanodisc membrane technology. The Campylobacter jejuni N-linked glycosylation pathway has been chosen as a model pathway in which all of the enzymes and substrates are biochemically accessible. We present the functional reconstitution of two enzymes responsible for the early membrane-committed steps in glycan assembly. Protein stoichiometry analysis, fluorescence-based approaches, and biochemical activity assays are used to demonstrate the colocalization of the two enzymes in nanodiscs. Isotopic labeling of the substrates reveals that undecaprenyl-phosphate is coincorporated into discs with the two enzymes, and furthermore, that both enzymes are functionally reconstituted and can sequentially convert the coembedded undecaprenyl-phosphate into undecaprenyl-diphosphate-linked disaccharide. These studies provide a proof-of-concept demonstrating that the nanodisc model membrane system represents a promising experimental platform for analyzing the multifaceted interactions among the enzymes involved in polyprenol-dependent glycan assembly pathways, the membrane-associated substrates, and the lipid bilayer. The stage is now set for exploration of the roles of the conserved polyprenols in promoting protein-protein interactions among pathway enzymes and processing of substrates through sequential steps in membrane-associated glycan assembly.

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Recent advances in nanodisc technology for membrane protein studies (2012–2017)

et al. 2017

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Historically, progress in membrane protein research has been hindered due to solubility issues. The introduction of biomembrane mimetics has since stimulated the field’s momentum. One mimetic, the nanodisc, has proved to be an exceptional system for solubilizing membrane proteins. Herein, we critically evaluate the advantages and imperfections from employing nanodiscs in biophysical and biochemical studies. Specifically, we examine the techniques that have been modified to study membrane proteins in nanodiscs. Techniques discussed include fluorescence microscopy, solution state/solid state NMR, electron microscopy, SAXS, and several mass spectroscopy methods. Newer techniques such as SPR, charge sensitive optical detection, and scintillation proximity assays are also reviewed. Lastly, we cover nanodiscs advancing nanotechnology through nanoplasmonic biosensing, lipoprotein-nanoplatelets, and sortase-mediated labeling of nanodiscs.

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Dynamic Interaction of the Escherichia coli Cell Division ZipA and FtsZ Proteins Evidenced in Nanodiscs

Cited by 45 publications

References 24 publications

Bacterial Division Proteins FtsZ and ZipA Induce Vesicle Shrinkage and Cell Membrane Invagination

Bacterial Division Proteins FtsZ and ZipA Induce Vesicle Shrinkage and Cell Membrane Invagination

Lipid bilayer nanodisc platform for investigating polyprenol-dependent enzyme interactions and activities

Recent advances in nanodisc technology for membrane protein studies (2012–2017)

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