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dc.contributor.authorTolchard, Jamesen
dc.contributor.authorWalpole, Samuel J.en
dc.contributor.authorMiles, Andrew J.en
dc.contributor.authorMaytum, Robinen
dc.contributor.authorEaglen, Lawrence A.en
dc.contributor.authorHackstadt, Teden
dc.contributor.authorWallace, B.A.en
dc.contributor.authorBlumenschein, Tharin M.A.en
dc.date.accessioned2018-09-06T09:13:59Z
dc.date.available2018-09-06T09:13:59Z
dc.date.issued2018-01-31
dc.identifier.citationTolchard J, Walpole SJ, Miles AJ, Maytum R, Eaglen LA, Hackstadt T, Wallace BA, Blumenschein TMA (2018) 'The intrinsically disordered Tarp protein from chlamydia binds actin with a partially preformed helix.', Scientific Reports, 31 (8), pp.1960-.en
dc.identifier.issn2045-2322
dc.identifier.pmid29386631
dc.identifier.doi10.1038/s41598-018-20290-8
dc.identifier.urihttp://hdl.handle.net/10547/622842
dc.description.abstractTarp (translocated actin recruiting phosphoprotein) is an effector protein common to all chlamydial species that functions to remodel the host-actin cytoskeleton during the initial stage of infection. In C. trachomatis, direct binding to actin monomers has been broadly mapped to a 100-residue region (726-825) which is predicted to be predominantly disordered, with the exception of a ~10-residue α-helical patch homologous to other WH2 actin-binding motifs. Biophysical investigations demonstrate that a Tarp726-825 construct behaves as a typical intrinsically disordered protein; within it, NMR relaxation measurements and chemical shift analysis identify the ten residue WH2-homologous region to exhibit partial α-helix formation. Isothermal titration calorimetry experiments on the same construct in the presence of monomeric G-actin show a well defined binding event with a 1:1 stoichiometry and Kd of 102 nM, whilst synchrotron radiation circular dichroism spectroscopy suggests the binding is concomitant with an increase in helical secondary structure. Furthermore, NMR experiments in the presence of G-actin indicate this interaction affects the proposed WH2-like α-helical region, supporting results from in silico docking calculations which suggest that, when folded, this α-helix binds within the actin hydrophobic cleft as seen for other actin-associated proteins.
dc.language.isoenen
dc.publisherNatureen
dc.relation.urlhttps://www.nature.com/articles/s41598-018-20290-8en
dc.rightsGreen - can archive pre-print and post-print or publisher's version/PDF
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/*
dc.subjectNMRen
dc.subjecttranslocated actin recruiting phosphoproteinen
dc.subjectTARPen
dc.subjectC521 Medical Microbiologyen
dc.titleThe intrinsically disordered Tarp protein from chlamydia binds actin with a partially preformed helixen
dc.typeArticleen
dc.identifier.journalScientific Reportsen
dc.identifier.pmcidPMC5792643
dc.date.updated2018-09-06T08:54:26Z
dc.description.noteopen access
html.description.abstractTarp (translocated actin recruiting phosphoprotein) is an effector protein common to all chlamydial species that functions to remodel the host-actin cytoskeleton during the initial stage of infection. In C. trachomatis, direct binding to actin monomers has been broadly mapped to a 100-residue region (726-825) which is predicted to be predominantly disordered, with the exception of a ~10-residue α-helical patch homologous to other WH2 actin-binding motifs. Biophysical investigations demonstrate that a Tarp726-825 construct behaves as a typical intrinsically disordered protein; within it, NMR relaxation measurements and chemical shift analysis identify the ten residue WH2-homologous region to exhibit partial α-helix formation. Isothermal titration calorimetry experiments on the same construct in the presence of monomeric G-actin show a well defined binding event with a 1:1 stoichiometry and Kd of 102 nM, whilst synchrotron radiation circular dichroism spectroscopy suggests the binding is concomitant with an increase in helical secondary structure. Furthermore, NMR experiments in the presence of G-actin indicate this interaction affects the proposed WH2-like α-helical region, supporting results from in silico docking calculations which suggest that, when folded, this α-helix binds within the actin hydrophobic cleft as seen for other actin-associated proteins.


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