Calcium Triggered Lα-H2 Phase Transition Monitored by Combined Rapid Mixing and Time-Resolved Synchrotron SAXS
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{"title"=>"Calcium triggered Lα-H2 phase transition monitored by combined rapid mixing and time-resolved synchrotron SAXS", "type"=>"journal", "authors"=>[{"first_name"=>"Anan", "last_name"=>"Yaghmur", "scopus_author_id"=>"6602560099"}, {"first_name"=>"Peter", "last_name"=>"Laggner", "scopus_author_id"=>"7005350700"}, {"first_name"=>"Barbara", "last_name"=>"Sartori", "scopus_author_id"=>"23969166400"}, {"first_name"=>"Michael", "last_name"=>"Rappolt", "scopus_author_id"=>"6701693796"}], "year"=>2008, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"44349098503", "pmid"=>"18446202", "isbn"=>"1932-6203", "pui"=>"352025025", "issn"=>"19326203", "scopus"=>"2-s2.0-44349098503", "doi"=>"10.1371/journal.pone.0002072"}, "id"=>"11cb5164-25f7-37e5-9a19-474a52fe0c6a", "abstract"=>"BACKGROUND: Awad et al. reported on the Ca(2+)-induced transitions of dioleoyl-phosphatidylglycerol (DOPG)/monoolein (MO) vesicles to bicontinuous cubic phases at equilibrium conditions. In the present study, the combination of rapid mixing and time-resolved synchrotron small-angle X-ray scattering (SAXS) was applied for the in-situ investigations of fast structural transitions of diluted DOPG/MO vesicles into well-ordered nanostructures by the addition of low concentrated Ca(2+) solutions.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: Under static conditions and the in absence of the divalent cations, the DOPG/MO system forms large vesicles composed of weakly correlated bilayers with a d-spacing of approximately 140 A (L(alpha)-phase). The utilization of a stopped-flow apparatus allowed mixing these DOPG/MO vesicles with a solution of Ca(2+) ions within 10 milliseconds (ms). In such a way the dynamics of negatively charged PG to divalent cation interactions, and the kinetics of the induced structural transitions were studied. Ca(2+) ions have a very strong impact on the lipidic nanostructures. Intriguingly, already at low salt concentrations (DOPG/Ca(2+)>2), Ca(2+) ions trigger the transformation from bilayers to monolayer nanotubes (inverted hexagonal phase, H(2)). Our results reveal that a binding ratio of 1 Ca(2+) per 8 DOPG is sufficient for the formation of the H(2) phase. At 50 degrees C a direct transition from the vesicles to the H(2) phase was observed, whereas at ambient temperature (20 degrees C) a short lived intermediate phase (possibly the cubic Pn3m phase) coexisting with the H(2) phase was detected.\\n\\nCONCLUSIONS/SIGNIFICANCE: The strong binding of the divalent cations to the negatively charged DOPG molecules enhances the negative spontaneous curvature of the monolayers and causes a rapid collapsing of the vesicles. The rapid loss of the bilayer stability and the reorganization of the lipid molecules within ms support the argument that the transition mechanism is based on a leaky fusion of the vesicles.", "link"=>"http://www.mendeley.com/research/calcium-triggered-l%CE%B1h2-phase-transition-monitored-combined-rapid-mixing-timeresolved-synchrotron-sax", "reader_count"=>36, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>11, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>2, "Student > Master"=>3, "Student > Bachelor"=>3, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>1, "Researcher"=>11, "Student > Doctoral Student"=>2, "Student > Ph. D. Student"=>9, "Student > Postgraduate"=>2, "Student > Master"=>3, "Student > Bachelor"=>3, "Lecturer"=>1, "Lecturer > Senior Lecturer"=>1, "Professor"=>2}, "reader_count_by_subject_area"=>{"Engineering"=>1, "Unspecified"=>1, "Environmental Science"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Materials Science"=>3, "Medicine and Dentistry"=>2, "Agricultural and Biological Sciences"=>2, "Pharmacology, Toxicology and Pharmaceutical Science"=>2, "Business, Management and Accounting"=>1, "Physics and Astronomy"=>8, "Chemistry"=>12}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Materials Science"=>{"Materials Science"=>3}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>2}, "Chemistry"=>{"Chemistry"=>12}, "Physics and Astronomy"=>{"Physics and Astronomy"=>8}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>2}, "Business, Management and Accounting"=>{"Business, Management and Accounting"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>1}, "Environmental Science"=>{"Environmental Science"=>1}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Japan"=>1, "Switzerland"=>1}, "group_count"=>1}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/933154"], "description"=>"<p>The investigated vesicles in the absence of Ca<sup>2+</sup> ions were composed of DOPG and MO at a constant molar ratio of 30∶70 and the total lipid content was 7 wt%. Eight different rapid-mixing investigations with different volume ratios of the vesicles (volume A) to the buffer (volume B) are summarized. In addition, the final Ca<sup>2+</sup> concentrations are given.</p>", "links"=>[], "tags"=>["molar", "ratios", "mixing", "vesicles", "pipes", "buffer", "containing", "68", "mm"], "article_id"=>603590, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.t002", "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_molar_ratios_of_DOPG_Ca_2_and_DOPG_MO_Ca_2_after_rapidly_mixing_the_DOPG_MO_based_vesicles_with_PIPES_buffer_pH_7_0_containing_68_mM_Ca_2_ions_/603590", "title"=>"The molar ratios of DOPG/[Ca<sup>2+</sup>] and (DOPG+MO)/[Ca<sup>2+</sup>] after rapidly mixing the DOPG/MO-based vesicles with PIPES buffer (pH 7.0) containing 68 mM Ca<sup>2+</sup> ions.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-21 10:37:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/932928"], "description"=>"<p>(A) The SAXS pattern is averaged from the data taken in the range of 100–400 ms after the rapid mixing. It indicates a possible formation of bicontinuous cubic phase of the symmetry Pn3m. The observed Bragg peaks and their presented <i>q</i>-values, suggest a coexistence with the H<sub>2</sub> phase (for details see text). (B) Temporal evolution of the observed lattice spacings.</p>", "links"=>[], "tags"=>["referring", "rapid-mixing"], "article_id"=>603371, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g008", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Intermediate_formation_referring_to_the_rapid_mixing_experiment_of_Figure_7_/603371", "title"=>"Intermediate formation referring to the rapid-mixing experiment of Figure 7.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:36:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/932197"], "description"=>"<p>In the stopped-flow apparatus, one syringe contained a buffer with Ca<sup>2+</sup> ions, whereas the other contained DOPG/MO-based vesicles. The rapid mixing was conducted within 10 ms and the formation of the inverse hexagonal phase (H<sub>2</sub>) was followed by millisecond time-resolved SAXS.</p>", "links"=>[], "tags"=>["stopped-flow", "synchrotron", "saxs"], "article_id"=>602638, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g001", "stats"=>{"downloads"=>5, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_of_the_combined_stopped_flow_and_synchrotron_SAXS_set_up_/602638", "title"=>"Schematic of the combined stopped-flow and synchrotron SAXS set-up.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:32:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/932705"], "description"=>"<p>All rapid-mixing experiments were carried out at 50°C. (A) The SAXS patterns of the DOPG/MO-based aqueous dispersions are displayed 71 s after the rapid mixing, i.e. approximately one minute after the turnovers were completed (<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002072#pone-0002072-g005\" target=\"_blank\">Figure 5</a>). It should be pointed out that for the two lowest salt concentrations the H<sub>2</sub> phase is coexisting with weakly correlated bilayers (the positions of the very weak first two diffraction orders are marked by arrows). (B) The <i>d<sub>10</sub></i>-spacing of the H<sub>2</sub> phase is displayed as a function of the final Ca<sup>2+</sup> concentration. The inset shows the <i>d</i>-spacing in dependence of the DOPG/Ca<sup>2+</sup> ratio. The electroneutral regime is highlighted in light grey.</p>", "links"=>[], "tags"=>["calcium-induced", "dependence"], "article_id"=>603151, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g006", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Comparison_of_the_calcium_induced_H_2_phase_in_dependence_of_the_final_salt_concentration_/603151", "title"=>"Comparison of the calcium-induced H<sub>2</sub> phase in dependence of the final salt concentration.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:35:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/932584"], "description"=>"<p>The solid red line shows the best single exponential fit to the data. The time constant, <i>k</i>, was determined to be 0.10±0.03 and 0.12±0.02 sec<sup>−1</sup> for panels (A) and (B), respectively.</p>", "links"=>[], "tags"=>["dependence", "formed", "referring"], "article_id"=>603025, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g005", "stats"=>{"downloads"=>1, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Time_dependence_of_the_intensity_A_and_the_d_spacing_B_of_the_first_order_reflection_of_the_rapidly_formed_H_2_phase_referring_to_the_experiment_given_in_Figure_4_/603025", "title"=>"Time dependence of the intensity (A), and the <i>d</i>-spacing (B) of the first order reflection of the rapidly formed H<sub>2</sub> phase referring to the experiment given in Figure 4.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:34:29"}
  • {"files"=>["https://ndownloader.figshare.com/files/932292"], "description"=>"<p>The investigated vesicles are composed of DOPG and MO at a molar ratio of 30∶70 with a total lipid content of 7 wt%. The best global fit to the experimental data is given by a solid red line (cp. data analysis). The inset shows the quality of the fit at higher <i>q</i>-values. The structural bilayer parameters are summarized in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002072#pone-0002072-t001\" target=\"_blank\">Table 1</a>.</p>", "links"=>[], "tags"=>["subtracted", "saxs", "vesicles"], "article_id"=>602738, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g002", "stats"=>{"downloads"=>2, "page_views"=>24, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Background_subtracted_SAXS_pattern_of_DOPG_MO_based_vesicles_at_25_C_in_the_absence_of_Ca_2_ions_/602738", "title"=>"Background subtracted SAXS pattern of DOPG/MO-based vesicles at 25°C in the absence of Ca<sup>2+</sup> ions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:32:46"}
  • {"files"=>["https://ndownloader.figshare.com/files/933184"], "description"=>"<p>The <i>d</i>-spacing, the headgroup position, <i>z<sub>H</sub></i>, the headgroup width, <i>σ<sub>H</sub></i>, and the width of the hydrophobic core, <i>σ<sub>C</sub></i>, are defined in <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002072#pone-0002072-g003\" target=\"_blank\"><b>Figure 3</b></a>. <i>ρ<sub>R</sub></i>, is the relative electron density of the bilayer trough set in relation to the headgroup density, <i>N<sub>mean</sub></i>, the membrane correlation number, <i>N<sub>diff</sub></i>, the fraction of diffuse scattering and <i>η</i>, the Caillé parameter.</p>", "links"=>[], "tags"=>["derived", "saxs", "vesicles"], "article_id"=>603630, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.t001", "stats"=>{"downloads"=>3, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fit_parameters_derived_from_the_GAP_evaluation_of_the_SAXS_pattern_Figure_2_for_the_DOPG_MO_based_vesicles_7_wt_lipid_T_50_C_/603630", "title"=>"Fit parameters derived from the GAP evaluation of the SAXS pattern (Figure 2) for the DOPG/MO-based vesicles (7 wt% lipid, T = 50°C).", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-21 10:38:01"}
  • {"files"=>["https://ndownloader.figshare.com/files/933066"], "description"=>"<p>On the left hand side, the classical vesicle fusion route is depicted. The formation of pores is widely believed to be the prerequisite for the formation of bicontinuous cubic (Q<sub>2</sub>) nanostructures <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002072#pone.0002072-Shearman1\" target=\"_blank\">[87]</a>, which upon further curvature frustration may transform into self-assembled monolayer tubes (H<sub>2</sub> phase). On the right hand side, the direct formation of an inverse lipid nanotube between two opposed bilayers is illustrated. For better understanding of the structural conversions, the headgroups of opposed monolayers are shown in light blue whereas the rest are depicted in blue (for further details see text).</p>", "links"=>[], "tags"=>["schematics", "pathways", "bilayer", "inverted", "monolayer"], "article_id"=>603510, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g009", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Two_schematics_of_the_proposed_pathways_from_the_bilayer_to_the_inverted_monolayer_tube_transition_/603510", "title"=>"Two schematics of the proposed pathways from the bilayer to the inverted monolayer tube transition.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:37:19"}
  • {"files"=>["https://ndownloader.figshare.com/files/932361"], "description"=>"<p>This model is composed of one Gaussian representing the electron density distribution of the polar headgroups at ±<i>z<sub>H</sub></i> and a second for the hydrophobic core with its centre at the bilayer mid-plane. The corresponding widths of the Gaussians are given by the standard deviations <i>σ<sub>H</sub></i> and <i>σ<sub>C</sub></i>, respectively. <i>d</i> defines the lattice spacing of the L<sub>α</sub> phase. The obtained results refer to the data of <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002072#pone-0002072-g002\" target=\"_blank\">Figure 2</a>.</p>", "links"=>[], "tags"=>["bilayer", "vesicles"], "article_id"=>602807, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g003", "stats"=>{"downloads"=>2, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Electron_density_bilayer_model_of_the_DOPG_MO_vesicles_at_25_C_in_the_absence_of_Ca_2_ions_/602807", "title"=>"Electron density bilayer model of the DOPG/MO vesicles at 25°C in the absence of Ca<sup>2+</sup> ions.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:33:11"}
  • {"files"=>["https://ndownloader.figshare.com/files/932815"], "description"=>"<p>The vesicle dispersion contained DOPG/MO with a molar ratio 30∶70 (7 wt% lipid), and the final salt concentration was 34 mM. The contour plot displays the first three reflections of the H<sub>2</sub> phase, but its formation is not immediate. In the first 400 ms, an intermediate phase is apparent. Two strong reflections are indicated by arrows and one weaker peak is circled by a dashed line.</p>", "links"=>[], "tags"=>["x-ray", "calcium", "induced"], "article_id"=>603257, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g007", "stats"=>{"downloads"=>3, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Time_resolved_X_ray_pattern_of_the_calcium_induced_H_2_phase_formation_at_20_176_C_/603257", "title"=>"Time-resolved X-ray pattern of the calcium induced H<sub>2</sub> phase formation at 20°C.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:35:53"}
  • {"files"=>["https://ndownloader.figshare.com/files/932465"], "description"=>"<p>The vesicle dispersion contained DOPG/MO with a molar ratio 30∶70 (7 wt% lipid), and the final salt concentration was 20 mM. The contour plot clearly displays the first three reflections of the H<sub>2</sub> phase; no indication for the formation of an intermediate phase is spotted.</p>", "links"=>[], "tags"=>["x-ray", "calcium-triggered"], "article_id"=>602907, "categories"=>["Immunology", "Biotechnology", "Chemistry", "Biophysics"], "users"=>["Anan Yaghmur", "Peter Laggner", "Barbara Sartori", "Michael Rappolt"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0002072.g004", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Time_resolved_X_ray_pattern_of_the_rapid_calcium_triggered_H_2_phase_formation_at_50_176_C_/602907", "title"=>"Time-resolved X-ray pattern of the rapid calcium-triggered H<sub>2</sub> phase formation at 50°C.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-21 10:33:47"}

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Relative Metric

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