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dc.date.accessioned2013-03-12T08:37:29Z
dc.date.available2013-03-12T08:37:29Z
dc.date.issued2008en_US
dc.date.submitted2008-11-18en_US
dc.identifier.citationHaugen, Mads Christofer. The three dimensional structure of plantaricin J. Masteroppgave, University of Oslo, 2008en_US
dc.identifier.urihttp://hdl.handle.net/10852/11401
dc.description.abstractThe bacteriocin plantaricin J/K consists of the two peptides PlnJ and PlnK, and the two peptides must be present for optimal bacteriocin function. In this study, the three dimensional structure of the PlnJ peptide was determined by use of NMR spectroscopy. 15N labelled PlnJ for structure determination was produced by Escherichia coli BL-21. A pGEV-2 vector, containing a gene encoding a fusion of the immunoglobulin binding domain of Streptococcal protein G (GB1 domain) and PlnJ, was used to transform the bacteria. Peptide production was achieved by inducing expression of the gene in 15N enriched minimal media (M9). The PlnJ target peptide was cleaved from the GB1 domain fusion partner with cyanogen bromide, and purified using reverse phase HPLC. Molecular weight was determined to be 2970, using a MALDI-TOF mass spectrometer, indicating a 99 % degree of 15N labeling. The produced PlnJ was found to be biologically active when mixed with equimolar amounts of synthetic PlnK prior to being exposed to cells sensitive to plantaricin J/K. Structural investigations of PlnJ were made using circular dichroism (CD) spectroscopy and NMR spectroscopy. Structuring was calculated from CD data using the absorbance at 222 nm as a quantitative measure of α-helical content, and by fitting the entire spectrum to the CD spectra of a data base using the CDPro software package. Structural constraints were obtained by analyzing NMR spectra: NOE distance constraints from a NOESY-HSQC spectrum and a NOESY spectrum, dihedral angle constraints calculated from the 3JHNHα found in a HNHα spectrum, φ and ψ torsion angle constraints obtained by matching chemical shifts to a data base using the TALOS software. These constraints were used to calculate a three dimensional structure using the CYANA software. All structural data supports an α-helical structure. Furthermore, the data suggests that the helical content is divided between two regions spanning amino acid residues 3-13 and 18-21 and that the molecule is linear. The side chains of the polar and the non polar amino acid residues of PlnJ are confined to separate sides of the long axis of the structure, making it an amphiphilic molecule.eng
dc.language.isoengen_US
dc.subjecttredimensjonal struktur antibakterielle peptider peptid antibiotikaen_US
dc.titleThe three dimensional structure of plantaricin J : Determination, by NMR spectroscopy, of the three dimensional structure of the plantaricin J component of the plantaricin J/K two-peptide bacteriocinen_US
dc.typeMaster thesisen_US
dc.date.updated2009-04-02en_US
dc.creator.authorHaugen, Mads Christoferen_US
dc.subject.nsiVDP::476en_US
dc.identifier.bibliographiccitationinfo:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft.au=Haugen, Mads Christofer&rft.title=The three dimensional structure of plantaricin J&rft.inst=University of Oslo&rft.date=2008&rft.degree=Masteroppgaveen_US
dc.identifier.urnURN:NBN:no-21770en_US
dc.type.documentMasteroppgaveen_US
dc.identifier.duo87037en_US
dc.contributor.supervisorPer Eugen Kristiansen, Jon Nissen-Meyer, Per Rogne og Gunnar Fimlanden_US
dc.identifier.bibsys091916623en_US
dc.identifier.fulltextFulltext https://www.duo.uio.no/bitstream/handle/10852/11401/2/oppgave.pdf


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