Liposome-Mediated Cellular Delivery of Active gp91phox
Publication Date
September 12, 2007
Journal
PLOS ONE
Authors
Bruno Marques, Lavinia Liguori, Marie Hélène Paclet, Ana Villegas Mendéz, et al
Volume
2
Issue
9
Pages
e856
DOI
http://doi.org/10.1371/journal.pone.0000856
Publisher URL
http://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0000856
PubMed
http://www.ncbi.nlm.nih.gov/pubmed/17848987
PubMed Central
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1955831
Europe PMC
http://europepmc.org/abstract/MED/17848987
Web of Science
000207455600003
Scopus
41149111854
Mendeley
http://www.mendeley.com/research/liposomemediated-cellular-delivery-active-gp91phox
Events
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Mendeley | Further Information

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Scopus | Further Information

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  • {"files"=>["https://ndownloader.figshare.com/files/945434"], "description"=>"<p>(A) Expression of membrane protein in presence of lipids analyzed by western blot using a monoclonal antibody anti-his. (B) Diaphorase assay of purified proteoliposomes gp91<sup>phox</sup> 221-C using NBT or INT as electron acceptor, n≥2. (C) Delivery gp91<sup>phox</sup> 221-C with proteoliposomes in HCT116 cells after 8 hours of incubation. As a negative control, cells were incubated with the same amount (0.5 µg) of empty liposomes. The proteins were detected by fluorescence microscopy using a monoclonal anti-his antibody. (D) Confocal analysis of the distribution of gp91<sup>phox</sup> 221-C protein delivered with proteoliposomes in HeLa cells after 8 hours of incubation. As a negative control, cells were incubated with 0.5 µg of empty liposomes or 0.5 µg of soluble gp91<sup>phox</sup> 221-C. Cells were labelled for the exogenous protein using an anti-gp91<sup>phox</sup> antibody (Red) and for the endogenous Clathrin (Green). The cells were examined under a Zeiss LSM510 (NLO) laser confocal microscope. The image plane was chosen to be near the equator of the cell body and nucleus. (E) Higher magnification of the white boxed area in (D) with the respective <i>xz</i> projections.</p>", "links"=>[], "tags"=>["221-c"], "article_id"=>615840, "categories"=>["Biochemistry", "Biotechnology", "Cell Biology"], "users"=>["Bruno Marques", "Lavinia Liguori", "Marie-Hélène Paclet", "Ana Villegas-Mendéz", "Romy Rothe", "Françoise Morel", "Jean-Luc Lenormand"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000856.g005", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_gp91_phox_221_C_proteoliposomes_/615840", "title"=>"gp91<sup>phox</sup> 221-C proteoliposomes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-09-12 01:37:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/465123"], "description"=>"<div><h3>Background</h3><p>Gp91<sup>phox</sup> is a transmembrane protein and the catalytic core of the NADPH oxidase complex of neutrophils. Lack of this protein causes chronic granulomatous disease (CGD), a rare genetic disorder characterized by severe and recurrent infections due to the incapacity of phagocytes to kill microorganisms.</p><h3>Methodology</h3><p>Here we optimize a prokaryotic cell-free expression system to produce integral mammalian membrane proteins.</p><h3>Conclusions</h3><p>Using this system, we over-express truncated forms of the gp91<sup>phox</sup> protein under soluble form in the presence of detergents or lipids resulting in active proteins with a “native-like” conformation. All the proteins exhibit diaphorase activity in the presence of cytosolic factors (p67<sup>phox</sup>, p47<sup>phox</sup>, p40<sup>phox</sup> and Rac) and arachidonic acid. We also produce proteoliposomes containing gp91<sup>phox</sup> protein and demonstrate that these proteins exhibit activities similar to their cellular counterpart. The proteoliposomes induce rapid cellular delivery and relocation of recombinant gp91<sup>phox</sup> proteins to the plasma membrane. Our data support the concept of cell-free expression technology for producing recombinant proteoliposomes and their use for functional and structural studies or protein therapy by complementing deficient cells in gp91<sup>phox</sup> protein.</p></div>", "links"=>[], "tags"=>["liposome-mediated", "cellular"], "article_id"=>151704, "categories"=>["Biochemistry", "Biotechnology", "Cell Biology"], "users"=>["Bruno Marques", "Lavinia Liguori", "Marie-Hélène Paclet", "Ana Villegas-Mendéz", "Romy Rothe", "Françoise Morel", "Jean-Luc Lenormand"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000856", "stats"=>{"downloads"=>0, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Liposome_Mediated_Cellular_Delivery_of_Active_gp91_phox_/151704", "title"=>"Liposome-Mediated Cellular Delivery of Active gp91<sup>phox</sup>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-09-12 00:28:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/944989"], "description"=>"<p>This schema summarizes the different prediction of topology for gp91<sup>phox</sup> by using different programs (Sosui, TMHMM, TMpred and TopPred) or the model published by Taylor <i>et al </i><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0000856#pone.0000856-Taylor2\" target=\"_blank\">[17]</a>.</p>", "links"=>[], "tags"=>["hydrophobic", "domains"], "article_id"=>615362, "categories"=>["Biochemistry", "Biotechnology", "Cell Biology"], "users"=>["Bruno Marques", "Lavinia Liguori", "Marie-Hélène Paclet", "Ana Villegas-Mendéz", "Romy Rothe", "Françoise Morel", "Jean-Luc Lenormand"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000856.g001", "stats"=>{"downloads"=>3, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Alignment_of_predicted_hydrophobic_domains_of_gp91_phox_/615362", "title"=>"Alignment of predicted hydrophobic domains of gp91<sup>phox</sup>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-09-12 01:29:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/945159"], "description"=>"<p>(A) Optimization of the solubility of the gp91<sup>phox</sup> truncated proteins by western blot using a monoclonal antibody anti-his. (B) Diaphorase assay of purified soluble truncated gp91<sup>phox</sup> proteins using NBT or INT as electron acceptor. Results were expressed as moles of NBT or INT reduced/min/mol gp91<sup>phox</sup> truncated forms. Results are expressed as the average of at least two experiments±SD.</p>", "links"=>[], "tags"=>["recombinant"], "article_id"=>615539, "categories"=>["Biochemistry", "Biotechnology", "Cell Biology"], "users"=>["Bruno Marques", "Lavinia Liguori", "Marie-Hélène Paclet", "Ana Villegas-Mendéz", "Romy Rothe", "Françoise Morel", "Jean-Luc Lenormand"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000856.g003", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Solubility_and_activity_of_gp91_phox_recombinant_proteins_/615539", "title"=>"Solubility and activity of gp91<sup>phox</sup> recombinant proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-09-12 01:32:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/945607"], "description"=>"<p>(A) Diaphorase assay of purified proteoliposomes gp91phox 90-N using NBT or INT as electron acceptor, n≥2. Cellular uptake of gp91<sup>phox</sup> 90-N proteins delivered with proteoliposomes in HeLa cells after 8 or 24 hours of incubation. The proteins were detected by fluorescence microscopy using a monoclonal anti-his antibody (B) or a monoclonal anti-gp91<sup>phox</sup> antibody (C).</p>", "links"=>[], "tags"=>["90-n"], "article_id"=>615976, "categories"=>["Biochemistry", "Biotechnology", "Cell Biology"], "users"=>["Bruno Marques", "Lavinia Liguori", "Marie-Hélène Paclet", "Ana Villegas-Mendéz", "Romy Rothe", "Françoise Morel", "Jean-Luc Lenormand"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000856.g006", "stats"=>{"downloads"=>2, "page_views"=>11, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_gp91_phox_90_N_proteoliposomes_/615976", "title"=>"gp91<sup>phox</sup> 90-N proteoliposomes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-09-12 01:39:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/945300"], "description"=>"<p>(A) Diaphorase activity measured with cytochrome <i>b</i><sub>558</sub> purified from neutrophils (10 pmol) in presence of cytosol from normal (N) or p67<sup>phox</sup> deficient (p67<sup>0</sup>) EBV B lymphocytes using NBT (black bars) or INT (open bars) as electron acceptor. (B) Diaphorase activity measured with the gp91<sup>phox</sup> 221-C protein (10 pmol) in presence of cytosol from normal (N) or p67<sup>phox</sup> deficient (p67<sup>0</sup>) EBV B lymphocytes using INT (open bars) as electron acceptor. Results are expressed as the mean activity of three experiments±SD. * indicates results significantly different (P<0.05) from the control performed in presence of normal cytosol.</p>", "links"=>[], "tags"=>["diaphorase"], "article_id"=>615676, "categories"=>["Biochemistry", "Biotechnology", "Cell Biology"], "users"=>["Bruno Marques", "Lavinia Liguori", "Marie-Hélène Paclet", "Ana Villegas-Mendéz", "Romy Rothe", "Françoise Morel", "Jean-Luc Lenormand"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000856.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Specificity_of_the_diaphorase_activity_measured_with_gp91_phox_recombinant_proteins_/615676", "title"=>"Specificity of the diaphorase activity measured with gp91<sup>phox </sup>recombinant proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-09-12 01:34:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/945057"], "description"=>"<p>(A) Schematic representation of the phagocyte NADPH oxydase and truncated forms gp91<sup>phox</sup> expressed in a bacterial cell-free translation system. Gp91<sup>phox</sup> full-length (FL) is presented at the top of the diagram: transmembrane domains are shown in black. FAD and NADPH binding-sites are represented in red and pink respectively. Truncated gp91<sup>phox</sup> were designed with a his-tag located either at the N or at the C terminal part of the protein. (B) Analysis of the expression of the ten gp91<sup>phox</sup> truncated proteins by western blot using a monoclonal antibody anti-his.</p>", "links"=>[], "tags"=>["truncated"], "article_id"=>615436, "categories"=>["Biochemistry", "Biotechnology", "Cell Biology"], "users"=>["Bruno Marques", "Lavinia Liguori", "Marie-Hélène Paclet", "Ana Villegas-Mendéz", "Romy Rothe", "Françoise Morel", "Jean-Luc Lenormand"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0000856.g002", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_truncated_gp91_phox_proteins_/615436", "title"=>"Expression of truncated gp91<sup>phox</sup> proteins.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2007-09-12 01:30:36"}

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  • {"unique-ip"=>"4", "full-text"=>"3", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"4"}
  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"4"}
  • {"unique-ip"=>"7", "full-text"=>"9", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"4", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"5"}
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  • {"unique-ip"=>"15", "full-text"=>"16", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"7"}
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  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"2"}
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  • {"unique-ip"=>"8", "full-text"=>"9", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"9"}
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  • {"unique-ip"=>"6", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"7"}
  • {"unique-ip"=>"7", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"8"}
  • {"unique-ip"=>"7", "full-text"=>"4", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"9"}
  • {"unique-ip"=>"6", "full-text"=>"3", "pdf"=>"4", "abstract"=>"1", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"10"}
  • {"unique-ip"=>"13", "full-text"=>"11", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"1"}
  • {"unique-ip"=>"11", "full-text"=>"11", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"11"}
  • {"unique-ip"=>"8", "full-text"=>"5", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2014", "month"=>"12"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"1"}
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  • {"unique-ip"=>"13", "full-text"=>"9", "pdf"=>"5", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"5", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2015", "month"=>"12"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"3"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"4"}
  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"2", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"5"}
  • {"unique-ip"=>"7", "full-text"=>"9", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"6"}
  • {"unique-ip"=>"4", "full-text"=>"5", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"7"}
  • {"unique-ip"=>"9", "full-text"=>"10", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"8"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"1", "cited-by"=>"0", "year"=>"2016", "month"=>"9"}
  • {"unique-ip"=>"5", "full-text"=>"3", "pdf"=>"3", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"10"}
  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"11"}
  • {"unique-ip"=>"5", "full-text"=>"5", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2016", "month"=>"12"}
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  • {"unique-ip"=>"3", "full-text"=>"3", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"3"}
  • {"unique-ip"=>"22", "full-text"=>"50", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"5"}
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  • {"unique-ip"=>"6", "full-text"=>"6", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"3", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"7"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"8"}
  • {"unique-ip"=>"3", "full-text"=>"2", "pdf"=>"2", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"10"}
  • {"unique-ip"=>"2", "full-text"=>"2", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"11"}
  • {"unique-ip"=>"6", "full-text"=>"7", "pdf"=>"0", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"1", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2017", "month"=>"12"}
  • {"unique-ip"=>"1", "full-text"=>"1", "pdf"=>"1", "abstract"=>"0", "scanned-summary"=>"0", "scanned-page-browse"=>"0", "figure"=>"0", "supp-data"=>"0", "cited-by"=>"0", "year"=>"2018", "month"=>"1"}
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Relative Metric

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