The Deletion of Several Amino Acid Stretches of Escherichia coli Alpha-Hemolysin (HlyA) Suggests That the Channel-Forming Domain Contains Beta-Strands
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{"title"=>"The deletion of several amino acid stretches of Escherichia coli alpha-hemolysin (HlyA) suggests that the channel-forming domain contains beta-strands", "type"=>"journal", "authors"=>[{"first_name"=>"Roland", "last_name"=>"Benz", "scopus_author_id"=>"7103013456"}, {"first_name"=>"Elke", "last_name"=>"Maier", "scopus_author_id"=>"7102488709"}, {"first_name"=>"Susanne", "last_name"=>"Bauer", "scopus_author_id"=>"57197976182"}, {"first_name"=>"Albrecht", "last_name"=>"Ludwig", "scopus_author_id"=>"7202500629"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"600627298", "sgr"=>"84914680484", "issn"=>"19326203", "pmid"=>"25463653", "scopus"=>"2-s2.0-84914680484", "doi"=>"10.1371/journal.pone.0112248"}, "id"=>"e7c98238-aecf-3c52-8cf3-d1dbc51ddfad", "abstract"=>"Escherichia coli α-hemolysin (HlyA) is a pore-forming protein of 110 kDa belonging to the family of RTX toxins. A hydrophobic region between the amino acid residues 238 and 410 in the N-terminal half of HlyA has previously been suggested to form hydrophobic and/or amphipathic α-helices and has been shown to be important for hemolytic activity and pore formation in biological and artificial membranes. The structure of the HlyA transmembrane channel is, however, largely unknown. For further investigation of the channel structure, we deleted in HlyA different stretches of amino acids that could form amphipathic β-strands according to secondary structure predictions (residues 71-110, 158-167, 180-203, and 264-286). These deletions resulted in HlyA mutants with strongly reduced hemolytic activity. Lipid bilayer measurements demonstrated that HlyAΔ71-110 and HlyAΔ264-286 formed channels with much smaller single-channel conductance than wildtype HlyA, whereas their channel-forming activity was virtually as high as that of the wildtype toxin. HlyAΔ158-167 and HlyAΔ180-203 were unable to form defined channels in lipid bilayers. Calculations based on the single-channel data indicated that the channels generated by HlyAΔ71-110 and HlyAΔ264-286 had a smaller size (diameter about 1.4 to 1.8 nm) than wildtype HlyA channels (diameter about 2.0 to 2.6 nm), suggesting that in these mutants part of the channel-forming domain was removed. Osmotic protection experiments with erythrocytes confirmed that HlyA, HlyAΔ71-110, and HlyAΔ264-286 form defined transmembrane pores and suggested channel diameters that largely agreed with those estimated from the single-channel data. Taken together, these results suggest that the channel-forming domain of HlyA might contain β-strands, possibly in addition to α-helical structures.", "link"=>"http://www.mendeley.com/research/deletion-several-amino-acid-stretches-escherichia-coli-alphahemolysin-hlya-suggests-channelforming-d", "reader_count"=>9, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>3, "Student > Master"=>2, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Researcher"=>1, "Student > Ph. D. Student"=>3, "Student > Master"=>2, "Other"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>2, "Agricultural and Biological Sciences"=>5, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>2}, "Unspecified"=>{"Unspecified"=>1}}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1813248"], "description"=>"<p>The single-channel conductance data of HlyA and HlyA<sub>Δ71–110</sub> were fitted by using the Renkin correction factor multiplied by the aqueous diffusion coefficients of the different cations. The single-channel conductance for the different cations taken from <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112248#pone-0112248-t002\" target=\"_blank\">Table<u> 2</u></a> was normalized to that observed for Rb<sup>+</sup> (hydrated ion radius  = 0.105 nm), which was set to 1.0, and plotted versus the hydrated ion radii taken from Table 3 of Maier <i>et al.</i><a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112248#pone.0112248-Maier1\" target=\"_blank\">[60]</a>. The points correspond to the single-channel conductance observed with Li<sup>+</sup>, Na<sup>+</sup>, K<sup>+</sup>, Cs<sup>+</sup>, N(CH<sub>3</sub>)<sub>4</sub><sup>+</sup>, N(C<sub>2</sub>H<sub>5</sub>)<sub>4</sub><sup>+</sup>, and Tris<sup>+</sup>, which were all used for the pore diameter estimation (see Discussion). (A) The fit (solid lines) is shown for wildtype HlyA channels with <i>r</i> = 1.6 nm (upper line) and <i>r</i> = 1.0 nm (lower line). The best fit was achieved with <i>r</i> = 1.3 nm (diameter = 2.6 nm), which corresponds to the broken line. (B) The fit (solid lines) is shown for the HlyA<sub>Δ71–110</sub> channels with <i>r</i> = 1.1 nm (upper line) and <i>r</i> = 0.7 nm (lower line). The best fit of all data was achieved with <i>r</i> = 0.9 nm (diameter = 1.8 nm), which corresponds to the broken line.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256929, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g007", "stats"=>{"downloads"=>0, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Calculation_of_the_channel_diameters_of_HlyA_and_HlyA_916_71_8211_110_from_the_single_channel_conductance_/1256929", "title"=>"Calculation of the channel diameters of HlyA and HlyA<sub>Δ71–110</sub> from the single-channel conductance.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813257"], "description"=>"<p>The single-channel conductance of the HlyA, HlyA<sub>Δ71–110</sub>, and HlyA<sub>Δ264–286</sub> channels is shown as a function of the KCl concentration in the aqueous phase. The solid lines represent the fit of the single-channel conductance data with eqn. <i>G(c) = G<sub>0</sub>⋅c<sub>0</sub><sup>+</sup></i> (a combination of eqs. (3–5 and 7) assuming the presence of negative point charges within the channel (for HlyA: 2.3 negative charges, <i>q = </i>−3.68×10<sup>–19</sup> As; for HlyA<sub>Δ71–110</sub>∶1.7 negative charges, <i>q</i> = -2.72×10<sup>–19</sup> As; for HlyA<sub>Δ264–286</sub>∶2 negative charges, <i>q = </i>-3.2×10<sup>–19</sup> As) and assuming a channel diameter of 2 nm, 1.4 nm, and 1.6 nm for HlyA, HlyA<sub>Δ71–110</sub>, and HlyA<sub>Δ264–286</sub>, respectively. <i>c</i>, concentration of the KCl solution in M (molar); <i>G</i>, average single-channel conductance in nS (nano Siemens, 10<sup>–9</sup> S); <i>G<sub>0</sub></i>, specific single-channel conductance in the absence of negative point charges given in pS/M. The broken, dotted, and fractured (straight) lines show the single-channel conductance of the HlyA, HlyA<sub>Δ264–286</sub>, and HlyA<sub>Δ71–110</sub> channels in the absence of point charges and correspond to linear functions between channel conductance and bulk aqueous concentration (eqn. (7); <i>G(c) = G<sub>0</sub>⋅c</i>).</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256936, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g008", "stats"=>{"downloads"=>2, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_charges_in_wildtype_and_mutant_HlyA_on_the_single_channel_conductance_/1256936", "title"=>"Effect of charges in wildtype and mutant HlyA on the single-channel conductance.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813260"], "description"=>"a<p>The 5′- and 3′-terminal halves of the primers represent the nucleotide sequences of <i>hlyA</i> flanking the desired deletions on both sides. The deletion site is indicated by a vertical bar (|).</p><p>Primers used for site-directed mutagenesis to study the effect of deletions of different stretches of amino acids of HlyA on hemolytic activity and properties of the HlyA channels.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256939, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.t001", "stats"=>{"downloads"=>6, "page_views"=>12, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Primers_used_for_site_directed_mutagenesis_to_study_the_effect_of_deletions_of_different_stretches_of_amino_acids_of_HlyA_on_hemolytic_activity_and_properties_of_the_HlyA_channels_/1256939", "title"=>"Primers used for site-directed mutagenesis to study the effect of deletions of different stretches of amino acids of HlyA on hemolytic activity and properties of the HlyA channels.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813264"], "description"=>"a<p>The membranes were formed from 1% (mass/volume) asolectin dissolved in n-decane. The aqueous solutions were unbuffered and had a pH of 6. The applied voltage was 20 mV, and the temperature was 20°C. The average single-channel conductance, <i>G</i> (i.e. current divided by voltage), was calculated from at least 80 single events. The standard deviation of the single-channel conductance was generally below ±15%. <i>c</i> is the concentration of the aqueous salt solutions. The single-channel conductance of wildtype HlyA of <i>E. coli</i> is given for comparison <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112248#pone.0112248-Benz3\" target=\"_blank\">[28]</a>. The values denoted with an asterix were measured during this study with purified HlyA. n.m. means not measured.</p><p>Average single-channel conductance, <i>G</i>, of HlyA<sub>Δ71–110</sub> and HlyA<sub>Δ264–286</sub> in different salt solutions.<sup>a.</sup></p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256942, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.t002", "stats"=>{"downloads"=>7, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Average_single_channel_conductance_G_of_HlyA_916_71_8211_110_and_HlyA_916_264_8211_286_in_different_salt_solutions_a_/1256942", "title"=>"Average single-channel conductance, <i>G</i>, of HlyA<sub>Δ71–110</sub> and HlyA<sub>Δ264–286</sub> in different salt solutions.<sup>a.</sup>", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813230"], "description"=>"<p>(A) SDS-PAGE of extracellular proteins from <i>E. coli</i> 5K containing different plasmids. Lane 1, molecular mass markers given in kDa; lane 2, <i>E. coli</i> 5K/pACYC184 (vector control); lane 3, <i>E. coli</i> 5K/pANN202–312* overproducing HlyA; lane 4, isogenic strain overproducing HlyA<sub>Δ71–110</sub>; lane 5, isogenic strain overproducing HlyA<sub>Δ158–167</sub>; lane 6, isogenic strain overproducing HlyA<sub>Δ180–203</sub>; lane 7, isogenic strain overproducing HlyA<sub>Δ264–286</sub>. The proteins in cell-free culture supernatants (harvested in the late log phase) were precipitated by addition of ice-cold trichloroacetic acid (final concentration, 10%), pelleted by centrifugation at 12,000×g, washed with acetone, dried under vacuum, and dissolved in sample buffer <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0112248#pone.0112248-Ludwig2\" target=\"_blank\">[11]</a>. Proteins from 100 µl culture supernatant were separated on the gel and visualized by silver staining. (B) Hemolytic phenotype of <i>E. coli</i> 5K/pANN202–312* overproducing HlyA and of isogenic strains overproducing the HlyA mutants with the indicated deletions. Bacteria from individual colonies were picked onto a sheep blood/Cm agar plate that was subsequently incubated for 24 hours at 37°C.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256918, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g001", "stats"=>{"downloads"=>3, "page_views"=>202, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Extracellular_secretion_and_hemolytic_activity_of_E_coli_HlyA_and_of_HlyA_mutants_/1256918", "title"=>"Extracellular secretion and hemolytic activity of <i>E. coli</i> HlyA and of HlyA mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813239"], "description"=>"<p>Wildtype and mutant HlyA were expressed in <i>E. coli</i> 5K/pANN202–312* and isogenic mutant strains, respectively, and purified from culture supernatants using preparative SDS-PAGE. Lane 1, molecular mass markers given in kDa; lane 2, HlyA; lane 3, HlyA<sub>Δ71–110</sub>; lane 4, HlyA<sub>Δ158–167</sub>; lane 5, HlyA<sub>Δ180–203</sub>; lane 6, HlyA<sub>Δ264–286</sub>. In each lane, 5 µg of protein was separated and visualized by Coomassie blue staining.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256920, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g002", "stats"=>{"downloads"=>1, "page_views"=>25, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SDS_PAGE_of_purified_E_coli_HlyA_and_of_HlyA_mutants_/1256920", "title"=>"SDS-PAGE of purified <i>E. coli</i> HlyA and of HlyA mutants.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813240"], "description"=>"<p>Single-channel recordings of asolectin membranes were performed in the presence of 50 ng/ml HlyA (left side, upper trace), 50 ng/ml HlyA<sub>Δ71–110</sub> (left side, lower trace), and 50 ng/ml HlyA<sub>Δ264–286</sub> (right side). The aqueous phase contained 150 mM KCl (pH 6). The applied membrane potential was 20 mV; T = 20°C. The average single-channel conductance was 520 pS for HlyA, 150 pS for HlyA<sub>Δ71–110</sub>, and 320 pS for HlyA<sub>Δ264–286</sub>.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256921, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g003", "stats"=>{"downloads"=>14, "page_views"=>18, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Single_channel_recordings_with_E_coli_HlyA_HlyA_916_71_8211_110_and_HlyA_916_264_8211_286_/1256921", "title"=>"Single-channel recordings with <i>E. coli</i> HlyA, HlyA<sub>Δ71–110</sub>, and HlyA<sub>Δ264–286</sub>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813241"], "description"=>"<p>Single-channel recordings of asolectin membranes were performed in the presence of 100 ng/ml HlyA<sub>Δ158–167</sub> (upper trace) and 150 ng/ml HlyA<sub>Δ180–203</sub> (lower trace). The aqueous phase contained 150 mM KCl (pH 6). The applied membrane potential was 20 mV; T = 20°C. The transient conductance steps in the upper trace (HlyA<sub>Δ158–167</sub>) had a conductance of about 600 to 700 pS. The mutant HlyA<sub>Δ180–203</sub> produced under the given conditions only current noise (fuzzy channels) and no defined conductance states.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256922, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g004", "stats"=>{"downloads"=>1, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Single_channel_recordings_with_HlyA_916_158_8211_167_and_HlyA_916_180_8211_203_/1256922", "title"=>"Single-channel recordings with HlyA<sub>Δ158–167</sub> and HlyA<sub>Δ180–203</sub>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813242"], "description"=>"<p>Sheep erythrocytes in saline solution (control) or in saline solution supplemented with 30 mM of different carbohydrates (arabinose, cellobiose, and melezitose, with diameters of 0.62, 0.92, and 1.14 nm, respectively) were incubated with the toxin at 37°C for 30 min (A) and 90 min (B). Erythrocyte lysis was determined as a function of increasing aerolysin concentrations.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256923, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g005", "stats"=>{"downloads"=>0, "page_views"=>16, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Results_of_osmotic_protection_experiments_with_aerolysin_of_A_sobria_/1256923", "title"=>"Results of osmotic protection experiments with aerolysin of <i>A. sobria</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/1813245"], "description"=>"<p>Sheep erythrocytes were incubated with the toxins at 37°C for 60 min in saline solution (control) or in saline solution supplemented with 30 mM of PEGs of different molecular masses (PEG 400, 600, 1000, 2000, 3000, and 4000, with diameters of 1.07, 1.32, 1.72, 2.47, 3.05, and 3.54 nm, respectively). The concentration of HlyA was 0.5 µg/ml and that of HlyA<sub>Δ71–110</sub> and HlyA<sub>Δ264–286</sub> 2.5 µg/ml. The degree of hemolysis is shown as a function of the molecular mass of the PEGs.</p>", "links"=>[], "tags"=>["110 kDa", "Several Amino", "pore formation", "channel structure", "structure predictions", "channel diameters", "acid residues 238", "lipid bilayers", "RTX toxins", "Osmotic protection experiments", "Lipid bilayer measurements", "mutants part", "HlyA transmembrane channel", "wildtype HlyA channels", "wildtype toxin", "1.8 nm", "HlyA mutants", "wildtype HlyA", "transmembrane pores"], "article_id"=>1256926, "categories"=>["Biological Sciences"], "users"=>["Roland Benz", "Elke Maier", "Susanne Bauer", "Albrecht Ludwig"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0112248.g006", "stats"=>{"downloads"=>0, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Results_of_osmotic_protection_experiments_with_HlyA_HlyA_916_71_8211_110_and_HlyA_916_264_8211_286_/1256926", "title"=>"Results of osmotic protection experiments with HlyA, HlyA<sub>Δ71–110</sub>, and HlyA<sub>Δ264–286</sub>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-12-02 03:20:41"}

PMC Usage Stats | Further Information

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