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dc.contributor.authorWommack, Andrew
dc.contributor.authorRobson, Scott A.
dc.contributor.authorWanniarachchi, Yoshitha A.
dc.contributor.authorWan, Andrea
dc.contributor.authorTurner, Christopher J.
dc.contributor.authorWagner, Gerhard
dc.contributor.authorNolan, Elizabeth Marie
dc.date.accessioned2013-12-10T20:08:37Z
dc.date.available2013-12-10T20:08:37Z
dc.date.issued2012-12
dc.date.submitted2012-11
dc.identifier.issn0006-2960
dc.identifier.issn1520-4995
dc.identifier.urihttp://hdl.handle.net/1721.1/82906
dc.description.abstractHuman defensin 5 (HD5) is a 32-residue host-defense peptide expressed in the gastrointestinal, reproductive, and urinary tracts that has antimicrobial activity. It exhibits six cysteine residues that are regiospecifically oxidized to form three disulfide bonds (Cys[superscript 3]–Cys[superscript 31], Cys[superscript 5]–Cys[superscript 20], and Cys[superscript 10]–Cys[superscript 30]) in the oxidized form (HD5[subscript ox]). To probe the solution structure and oligomerization properties of HD5[subscript ox], and select mutant peptides lacking one or more disulfide bonds, NMR solution studies and analytical ultracentrifugation experiments are reported in addition to in vitro peptide stability assays. The NMR solution structure of HD5[subscript ox], solved at pH 4.0 in 90:10 H2O/D2O, is presented (PDB: 2LXZ). Relaxation T[subscript 1]/T[subscript 2] measurements and the rotational correlation time (τc) estimated from a [superscript 15]N-TRACT experiment demonstrate that HD5[subscript ox] is dimeric under these experimental conditions. Exchange broadening of the Hα signals in the NMR spectra suggests that residues 19–21 (Val[superscript 19]–Cys[superscript 20]–Glu[superscript 21]) contribute to the dimer interface in solution. Exchange broadening is also observed for residues 7–14 comprising the loop. Sedimentation velocity and equilibrium studies conducted in buffered aqueous solution reveal that the oligomerization state of HD5[subscript ox] is pH-dependent. Sedimentation coefficients of ca. 1.8 S and a molecular weight of 14 363 Da were determined for HD5[subscript ox] at pH 7.0, supporting a tetrameric form ([HD5ox] ≥ 30 μM). At pH 2.0, a sedimentation coefficient of ca. 1.0 S and a molecular weight of 7079 Da, corresponding to a HD5ox dimer, were obtained. Millimolar concentrations of NaCl, CaCl[subscript 2], and MgCl[subscript 2] have a negligible effect on the HD5[subscript ox] sedimentation coefficients in buffered aqueous solution at neutral pH. Removal of a single disulfide bond results in a loss of peptide fold and quaternary structure. These biophysical investigations highlight the dynamic and environmentally sensitive behavior of HD5[subscript ox] in solution, and provide important insights into HD5[subscript ox] structure/activity relationships and the requirements for antimicrobial action.en_US
dc.description.sponsorshipNational Institutes of Health (U.S.) (NIH Grant DP2OD007045)en_US
dc.description.sponsorshipNational Institute of General Medical Sciences (U.S.) (NIH Grant P01 GM047467)en_US
dc.description.sponsorshipNational Institute for Biomedical Imaging and Bioengineering (U.S.)en_US
dc.description.sponsorshipNational Institute for Biomedical Imaging and Bioengineering (U.S.) (NIH Grant EB-002026)en_US
dc.description.sponsorshipMassachusetts Institute of Technology. Dept. of Chemistryen_US
dc.language.isoen_US
dc.publisherAmerican Chemical Societyen_US
dc.relation.isversionofhttp://dx.doi.org/10.1021/bi301255uen_US
dc.rightsArticle is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use.en_US
dc.sourcePMCen_US
dc.titleNMR Solution Structure and Condition-Dependent Oligomerization of the Antimicrobial Peptide Human Defensin 5en_US
dc.typeArticleen_US
dc.identifier.citationWommack, Andrew J., Scott A. Robson, Yoshitha A. Wanniarachchi, Andrea Wan, Christopher J. Turner, Gerhard Wagner, and Elizabeth M. Nolan. “NMR Solution Structure and Condition-Dependent Oligomerization of the Antimicrobial Peptide Human Defensin 5.” Biochemistry 51, no. 48 (December 4, 2012): 9624-9637.en_US
dc.contributor.departmentMassachusetts Institute of Technology. Department of Chemistryen_US
dc.contributor.departmentFrancis Bitter Magnet Laboratory (Massachusetts Institute of Technology)en_US
dc.contributor.mitauthorWommack, Andrewen_US
dc.contributor.mitauthorWanniarachchi, Yoshitha A.en_US
dc.contributor.mitauthorWan, Andreaen_US
dc.contributor.mitauthorTurner, Christopher J.en_US
dc.contributor.mitauthorNolan, Elizabeth M.en_US
dc.relation.journalBiochemistryen_US
dc.eprint.versionAuthor's final manuscripten_US
dc.type.urihttp://purl.org/eprint/type/JournalArticleen_US
eprint.statushttp://purl.org/eprint/status/PeerRevieweden_US
dspace.orderedauthorsWommack, Andrew J.; Robson, Scott A.; Wanniarachchi, Yoshitha A.; Wan, Andrea; Turner, Christopher J.; Wagner, Gerhard; Nolan, Elizabeth M.en_US
dc.identifier.orcidhttps://orcid.org/0000-0002-6153-8803
mit.licensePUBLISHER_POLICYen_US


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