Surfactosomes: a novel approach to the reconstitution and 2‐D crystallisation of membrane proteins

Reviakine, Ilya; Stoylova, Svetla; Holzenburg, Andreas

doi:10.1016/0014-5793(96)00063-4

Cited by 5 publications

(3 citation statements)

References 27 publications

(35 reference statements)

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“…Such a stoichiometry suggests that the active unit of V o is a dimer of subunit c and, in fact, subunit c does form stable dimers in SDS [211]. A recent analysis of twodimensional arrays of subunit c in detergent also suggested that the unit cell is most likely a dimer of subunit c [212].…”

Section: H + /Atp Stoichiometrymentioning

confidence: 98%

The vacuolar H+-ATPase: a universal proton pump of eukaryotes

Finbow

Harrison

1997

Biochemical Journal

239

179

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The vacuolar H + -ATPase (V-ATPase) is a universal component of eukaryotic organisms. It is present in the membranes of many organelles, where its proton-pumping action creates the low intravacuolar pH found, for example, in lysosomes. In addition, there are a number of differentiated cell types that have V-ATPases on their surface that contribute to the physiological functions of these cells. The V-ATPase is a multi-subunit enzyme composed of a membrane sector and a cytosolic catalytic sector. It is related to the familiar F o F " ATP synthase (F-ATPase), having the same basic architectural construction, and many of the subunits from

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Section: H + /Atp Stoichiometrymentioning

confidence: 98%

The vacuolar H+-ATPase: a universal proton pump of eukaryotes

Finbow

Harrison

1997

Biochemical Journal

239

179

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“…In this respect, pentadecafluorooctanoate (PFO), or perfluorooctanoate, has been applied to purify, solubilize and maintain a membrane protein with quaternary structure, and to act as an alternative in monitoring oligomerization of some membrane proteins or peptide molecules by PAGE [11][12][13][14][15]. The membrane proteins tested with PFO include: the cystic fibrosis TM conductance regulator (CFTR) [16,17], the vanilloid receptor subtype 1 demonstrated to be predominantly tetrameric [18], the human xeroderma pigmentosum group A protein shown as disulfide bond-independent dimer [19], and the self-associated TM of Na,K-ATPase gsubunit [20].…”

Section: Introductionmentioning

confidence: 99%

The application of perfluorooctanoate to investigate trimerization of the human immunodeficiency virus‐1 gp41 ectodomain by electrophoresis

et al. 2008

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The transmembrane glycoprotein gp41 of human immunodeficiency virus has been proposed to form trimer-of-hairpin during virus-cell membrane fusion. To investigate its oligomerization propensity under soluble and membrane-mimic conditions, sodium salt of perfluorooctanoate (PFO) was applied. A recombinant gp41 ectodomain devoid of disulfide linkage was overexpressed in Escherichia coli and characterized by MS and circular dichroism spectropolarimetry in PFO solution in comparison to SDS. The helical content of this ectodomain in PFO is higher than that in SDS. Notably, PFO employed in PAGE clearly conduced to the formation of trimer under the optimized condition as visualized in the gel. In addition, the proteins expressed from the two mutants in the heptad repeat (HR) domains of gp41, I62P, and N126K, were also examined by the PFO-PAGE analysis for functional ramification of molecular organization. Remarkably, the I62P mutation completely abolished the gp41 trimer formation, whereas the N126K mutation resulted in a more stable trimer. The data suggested that PFO-PAGE analysis is appropriate for evaluating the effect of mutations on the trimerization of gp41 and other fusion proteins which may be implicated in the alteration of their fusogenicity.

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“…Fluorinated surfactants can prevent aggregation of membrane proteins in solution and chaperone their insertion into the membranes without partitioning themselves into lipid bilayers (Rodnin et al 2008; Kyrychenko et al 2012). Interestingly, Holzenburg and colleagues used ammonium perfluoroonate to solubilize the V-type H+-ATPase for 2D crystallization (Reviakine et al 1996). In a most recent paper, Raunser and colleagues (Efremov et al 2014) obtained a 6.1 Å resolution of the ryanodine receptor that is clearly inserted into nanodiscs.…”

Section: Future Prospectsmentioning

confidence: 99%

Following Natures Lead: On the Construction of Membrane-Inserted Toxins in Lipid Bilayer Nanodiscs

et al. 2015

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Bacterial toxin or viral entry into the cell often requires cell surface binding and endocytosis. The endosomal acidification induces a limited unfolding/refolding and membrane insertion reaction of the soluble toxins or viral proteins into their translocation competent or membrane inserted states. At the molecular level, the specific orientation and immobilization of the pre-transitioned toxin on the cell surface is often an important prerequisite prior to cell entry. We propose that structures of some toxin membrane insertion complexes may be observed through procedures where one rationally immobilizes the soluble toxin so that potential unfolding ↔ refolding transitions that occur prior to membrane insertion orientate away from the immobilization surface in the presence of lipid micelle pre-nanodisc structures. As a specific example, the immobilized prepore form of the anthrax toxin pore translocon or protective antigen can be transitioned, inserted into a model lipid membrane (nanodiscs), and released from the immobilized support in its membrane solubilized form. This particular strategy, although unconventional, is a useful procedure for generating pure membrane-inserted toxins in nanodiscs for electron microscopy structural analysis. In addition, generating a similar immobilized platform on label-free biosensor surfaces allows one to observe the kinetics of these acid-induced membrane insertion transitions. These platforms can facilitate the rational design of inhibitors that specifically target the toxin membrane insertion transitions that occur during endosomal acidification. This approach may lead to a new class of direct anti-toxin inhibitors.

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Surfactosomes: a novel approach to the reconstitution and 2‐D crystallisation of membrane proteins

Cited by 5 publications

References 27 publications

The vacuolar H+-ATPase: a universal proton pump of eukaryotes

The vacuolar H+-ATPase: a universal proton pump of eukaryotes

The application of perfluorooctanoate to investigate trimerization of the human immunodeficiency virus‐1 gp41 ectodomain by electrophoresis

Following Natures Lead: On the Construction of Membrane-Inserted Toxins in Lipid Bilayer Nanodiscs

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