DipA, a Pore-Forming Protein in the Outer Membrane of Lyme Disease Spirochetes Exhibits Specificity for the Permeation of Dicarboxylates
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{"title"=>"DipA, a pore-forming protein in the outer membrane of lyme disease spirochetes exhibits specificity for the permeation of dicarboxylates", "type"=>"journal", "authors"=>[{"first_name"=>"Marcus", "last_name"=>"Thein", "scopus_author_id"=>"15726731500"}, {"first_name"=>"Mari", "last_name"=>"Bonde", "scopus_author_id"=>"36799501600"}, {"first_name"=>"Ignas", "last_name"=>"Bunikis", "scopus_author_id"=>"23968838800"}, {"first_name"=>"Katrin", "last_name"=>"Denker", "scopus_author_id"=>"8707079600"}, {"first_name"=>"Albert", "last_name"=>"Sickmann", "scopus_author_id"=>"7004512650"}, {"first_name"=>"Sven", "last_name"=>"Bergström", "scopus_author_id"=>"35594805100"}, {"first_name"=>"Roland", "last_name"=>"Benz", "scopus_author_id"=>"7103013456"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84860999626", "doi"=>"10.1371/journal.pone.0036523", "pui"=>"364802799", "pmid"=>"22590556", "scopus"=>"2-s2.0-84860999626", "issn"=>"19326203", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)"}, "id"=>"6aed53bb-f776-39da-86cd-e82f5e172989", "abstract"=>"Lyme disease Borreliae are highly dependent on the uptake of nutrients provided by their hosts. Our study describes the identification of a 36 kDa protein that functions as putative dicarboxylate-specific porin in the outer membrane of Lyme disease Borrelia. The protein was purified by hydroxyapatite chromatography from Borrelia burgdorferi B31 and designated as DipA, for dicarboxylate-specific porin A. DipA was partially sequenced, and corresponding genes were identified in the genomes of B. burgdorferi B31, Borrelia garinii PBi and Borrelia afzelii PKo. DipA exhibits high homology to the Oms38 porins of relapsing fever Borreliae. B. burgdorferi DipA was characterized using the black lipid bilayer assay. The protein has a single-channel conductance of 50 pS in 1 M KCl, is slightly selective for anions with a permeability ratio for cations over anions of 0.57 in KCl and is not voltage-dependent. The channel could be partly blocked by different di- and tricarboxylic anions. Particular high stability constants up to about 28,000 l/mol (in 0.1 M KCl) were obtained among the 11 tested anions for oxaloacetate, 2-oxoglutarate and citrate. The results imply that DipA forms a porin specific for dicarboxylates which may play an important role for the uptake of specific nutrients in different Borrelia species.", "link"=>"http://www.mendeley.com/research/dipa-poreforming-protein-outer-membrane-lyme-disease-spirochetes-exhibits-specificity-permeation-dic", "reader_count"=>14, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Librarian"=>1, "Researcher"=>3, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Librarian"=>1, "Researcher"=>3, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Other"=>1, "Student > Bachelor"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>4, "Philosophy"=>1, "Physics and Astronomy"=>3, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Physics and Astronomy"=>{"Physics and Astronomy"=>3}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>4}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>2}, "Philosophy"=>{"Philosophy"=>1}}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/641231"], "description"=>"<p>Ultrathin cryosections were prepared from the <i>B. burgdorferi</i> Osp-less mutant B313 at −110°C, embedded in gelatin. The immungold particles were visualized by sequential incubation of the fixed cells by the polyclonal rabbit antibodies detecting (A) DipA or (B) pre-serum and the anti-rabbit antibodies GAR10.</p>", "links"=>[], "tags"=>["electron", "micrograph", "immunogold", "labeled", "dipa", "membrane", "osp-less", "mutant"], "article_id"=>311710, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.g004", "stats"=>{"downloads"=>3, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transmission_electron_micrograph_showing_immunogold_labeled_DipA_in_the_outer_membrane_of_the_B_burgdorferi_Osp_less_mutant_B313_/311710", "title"=>"Transmission electron micrograph showing immunogold labeled DipA in the outer membrane of the <i>B. burgdorferi</i> Osp-less mutant B313.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:25:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/640903"], "description"=>"<p>Approximately 1–10 ng of outer membrane fraction (OMF) of <i>B. burgdorferi</i> B31 Δp66 or hydroxyapatite chromatography-purified (HAC) DipA was separated by 12% SDS-PAGE and silver-stained. The positions of molecular mass standards in kDa are shown at the left panel.</p>", "links"=>[], "tags"=>["purified"], "article_id"=>311386, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.g001", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Analysis_of_purified_DipA_/311386", "title"=>"Analysis of purified DipA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:24:06"}
  • {"files"=>["https://ndownloader.figshare.com/files/641739"], "description"=>"<p><i>V</i><sub>m</sub> is defined as the difference between the potential at the dilute side (100 mM) and the potential at the concentrated side (500 mM). The aqueous electrolyte solutions were buffered with 10 mM Tris-HCl (pH 7.5); temperature  = 20°C; The permeability ratio <i>P</i><sub>c</sub>/<i>P</i><sub>a</sub> was calculated using the Goldman-Hodgin-Katz equation <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036523#pone.0036523-Benz5\" target=\"_blank\">[62]</a> from at least three individual experiments.</p>", "links"=>[], "tags"=>["membrane", "potentials", "diphytanoyl", "membranes", "dipa", "five-fold", "gradient"], "article_id"=>312218, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.t002", "stats"=>{"downloads"=>2, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Zero_current_membrane_potentials_V_m_of_diphytanoyl_phosphatidylcholine_n_decane_membranes_in_the_presence_of_DipA_measured_for_a_five_fold_concentration_gradient_of_different_electrolytes_/312218", "title"=>"Zero-current membrane potentials (<i>V</i><sub>m</sub>) of diphytanoyl phosphatidylcholine/<i>n</i>-decane membranes in the presence of DipA measured for a five-fold concentration gradient of different electrolytes.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:28:24"}
  • {"files"=>["https://ndownloader.figshare.com/files/641463"], "description"=>"<p>The membrane was formed from diphytanoyl phosphatidylcholine/<i>n</i>-decane. The aqueous phase contained ∼100 ng ml<sup>−1</sup> DipA protein, 0.1 M KCl, 10 mM Tris-Cl, pH 7.5 and dicarboxylates at concentrations as indicated; temperature  = 20°C; voltage  = 20 mV.</p>", "links"=>[], "tags"=>["membrane", "conductance", "induced", "dipa", "malate", "2-oxoglutarate", "phthalate"], "article_id"=>311944, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.g006", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Titration_of_membrane_conductance_induced_by_DipA_with_malate_A_2_oxoglutarate_B_and_phthalate_C_/311944", "title"=>"Titration of membrane conductance induced by DipA with malate (A), 2-oxoglutarate (B) and phthalate (C).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:26:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/641151"], "description"=>"<p>Immunoblot analysis with antiserum against <i>B. burgdorferi</i> DipA resulted in clear signals of DipA and its homologues in the total protein fractions (TP) of the Lyme disease agents <i>B. burgdorferi</i> (<i>B. b.</i>), <i>B. afzelii</i> (<i>B. a.</i>), <i>B. garinii</i> (<i>B. g.</i>) and the relapsing fever agents <i>B. crocidurae</i> (<i>B. c.</i>), <i>B. duttonii</i> (<i>B. d.</i>), <i>B. hermsii</i> (<i>B. h.</i>), <i>B. hispanica</i> (<i>B. his</i>) and <i>B. recurrentis</i> (<i>B. r.</i>). The immunoblot signal of the hydroxyapatite chromatography (HAC) purified <i>B. burgdorferi</i> (<i>B. b.</i>) DipA is on the right. The position of molecular mass standard in kDa is shown at the left.</p>", "links"=>[], "tags"=>["dipa", "lyme", "relapsing"], "article_id"=>311637, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.g003", "stats"=>{"downloads"=>2, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Detection_of_DipA_in_Lyme_disease_and_relapsing_fever_spirochetes_/311637", "title"=>"Detection of DipA in Lyme disease and relapsing fever spirochetes.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:25:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/641330"], "description"=>"<p>(A) Single-channel conductance observed for DipA in a diphytanoyl phosphatidylcholine/<i>n</i>-decane (PC) membrane. About 10 ng ml<sup>−1</sup> of purified DipA of about was added to a PC lipid bilayer, bathed in 1 M KCl. (B) Histogram of individual single-channel events observed for purified DipA. The average single-channel conductance was 50 pS for a total number of 140 single steps; temperature  = 20°C; voltage  = 20 mV.</p>", "links"=>[], "tags"=>["Infectious diseases", "microbiology", "biotechnology", "biophysics", "Biochemistry"], "article_id"=>311814, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.g005", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pore_forming_activity_of_DipA_/311814", "title"=>"Pore-forming activity of DipA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:26:21"}
  • {"files"=>["https://ndownloader.figshare.com/files/641692"], "description"=>"<p>The organic anions are important key metabolites of <i>Borrelia</i> species. The membranes were formed from diphytanoyl phosphatidylcholine dissolved in <i>n</i>-decane. The buffered aqueous phase (1 mM Tris-HCl pH 7.5) contained purified DipA in a concentration of about 100 ng/ml and 0.1 M KCl; temperature  = 20°C; voltage  = 20 mV. The stability constants were derived from titration experiments similar to those shown in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036523#pone-0036523-g003\" target=\"_blank\">Fig. 3</a>. The stability constant, <i>K</i>, is given as the mean of <i>n</i> experiments ± SD. <i>K</i><sub>s</sub> is the half-saturation constant.</p>", "links"=>[], "tags"=>["binding", "anions", "dipa"], "article_id"=>312180, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.t003", "stats"=>{"downloads"=>4, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Stability_constants_K_for_the_binding_of_different_organic_anions_to_the_DipA_channel_/312180", "title"=>"Stability constants, <i>K</i>, for the binding of different organic anions to the DipA channel.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:28:12"}
  • {"files"=>["https://ndownloader.figshare.com/files/641030"], "description"=>"<p>The alignment was performed using Pole Bioinformatique Lyonnaise Network Protein Sequence Analysis (<a href=\"http://npsa-pbil.ibcp.fr\" target=\"_blank\">http://npsa-pbil.ibcp.fr</a>). Amino acids identical in all four proteins are highlighted in red, strongly similar amino acids (:) are given in green and weakly similar ones (.) in blue. The putative beta strands in DipA of <i>B. burgdorferi</i> are indicated by blue bars as derived from secondary structure prediction programs <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036523#pone.0036523-Bagos1\" target=\"_blank\">[55]</a>, <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036523#pone.0036523-Gromiha1\" target=\"_blank\">[56]</a>. The cleavage site of the N-terminal signal peptide of DipA of <i>B. burgdorferi</i> B31 as predicted by the program SignalP 3.0 (<a href=\"http://www.cbs.dtu.dk/services/SignalP/\" target=\"_blank\">http://www.cbs.dtu.dk/services/SignalP/</a>) with maximum probability (about 50%) is marked by a black bar <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036523#pone.0036523-Bendtsen1\" target=\"_blank\">[54]</a>. This is the same site as has been found previously for the N-terminal end of Oms38 of <i>B. duttonii</i> by N-terminal sequencing <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036523#pone.0036523-Thein1\" target=\"_blank\">[48]</a>. Partial peptide sequences obtained by mass spectrometry are in bold and highlighted in yellow.</p>", "links"=>[], "tags"=>["alignment", "dipa", "b31", "pbi", "pko", "duttonii"], "article_id"=>311505, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.g002", "stats"=>{"downloads"=>2, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Amino_acid_sequence_alignment_of_DipA_of_B_burgdorferi_B31_B_b_B_garinii_PBi_B_g_B_afzelii_PKo_B_a_and_B_duttonii_B_d_/311505", "title"=>"Amino acid sequence alignment of DipA of B. <i>burgdorferi</i> B31 (B.b.), <i>B. garinii</i> PBi (B.g.) <i>B. afzelii</i> PKo (B.a.) and B. duttonii (B.d.).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:24:44"}
  • {"files"=>["https://ndownloader.figshare.com/files/641587"], "description"=>"<p>The data were taken from <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036523#pone-0036523-g006\" target=\"_blank\">figure 6</a> and analyzed using equation 4. The straight lines correspond to stability constants <i>K</i>, for malate binding of 926 l/mol (<i>K</i><sub>s</sub> = 1.08 mM), for 2-oxoglutarate 20,298 l/mol (<i>K</i><sub>s</sub> = 0.05 mM) and for phthalate 5,152 l/mol (<i>K</i><sub>s</sub> = 0.19 mM).</p>", "links"=>[], "tags"=>["plots", "inhibition", "membrane", "conductance", "malate", "2-oxoglutarate", "phthalate"], "article_id"=>312066, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.g007", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Lineweaver_Burk_plots_of_the_inhibition_of_membrane_conductance_by_malate_A_2_oxoglutarate_B_and_phthalate_C_/312066", "title"=>"Lineweaver-Burk plots of the inhibition of membrane conductance by malate (A), 2-oxoglutarate (B) and phthalate (C).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-20 05:27:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/641789"], "description"=>"<p>The membranes were formed from diphytanoyl phosphatidylcholine dissolved in <i>n</i>-decane. The aqueous electrolyte solutions were unbuffered and had a pH of ∼6 unless otherwise indicated; temperature  = 20°C; voltage  = 20 mV. The average single-channel conductance, <i>G</i>, was calculated from at least 30 single reconstitution events of DipA.</p>", "links"=>[], "tags"=>["single-channel", "conductance", "dipa", "electrolyte"], "article_id"=>312270, "categories"=>["Microbiology", "Biotechnology", "Biochemistry", "Infectious Diseases", "Biophysics"], "users"=>["Marcus Thein", "Mari Bonde", "Ignas Bunikis", "Katrin Denker", "Albert Sickmann", "Sven Bergström", "Roland Benz"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0036523.t001", "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Average_single_channel_conductance_G_of_DipA_in_different_electrolyte_solutions_/312270", "title"=>"Average single-channel conductance (<i>G</i>) of DipA in different electrolyte solutions.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-20 05:28:38"}

PMC Usage Stats | Further Information

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Relative Metric

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