Crystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocol

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dc.contributor.authorvan Rooyen, J M
dc.contributor.authorAbratt, V R
dc.contributor.authorBelrhali, H
dc.contributor.authorSewell, B T
dc.date.accessioned2016-07-25T10:48:09Z
dc.date.available2016-07-25T10:48:09Z
dc.date.issued2010
dc.date.updated2016-07-12T14:34:24Z
dc.description.abstractGlutamine synthetase enzymes (GSs) are large oligomeric enzymes that play a critical role in nitrogen metabolism in all forms of life. To date, no crystal structures exist for the family of large (1 MDa) type III GS enzymes, which only share 9% sequence identity with the well characterized GSI and GSII enzymes. Here we present a novel protocol for the isolation of untagged Bacteroides fragilis GlnN expressed in an auxotrophic Escherichia coli strain. The rapid and scalable two-step protocol utilized differential precipitation by divalent cations followed by affinity chromatography to produce suitable quantities of homogenous material for structural characterization. Subsequent optimizations to the sample stability and solubility led to the discovery of conditions for the production of the first diffraction quality crystals of a type III GS enzyme.en_ZA
dc.identifierhttp://dx.doi.org/10.1016/j.pep.2010.06.007
dc.identifier.apacitationvan Rooyen, J. M., Abratt, V. R., Belrhali, H., & Sewell, B. T. (2010). Crystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocol. <i>Protein Expression and Purification</i>, http://hdl.handle.net/11427/20674en_ZA
dc.identifier.chicagocitationvan Rooyen, J M, V R Abratt, H Belrhali, and B T Sewell "Crystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocol." <i>Protein Expression and Purification</i> (2010) http://hdl.handle.net/11427/20674en_ZA
dc.identifier.citationVan Rooyen, J. M., Abratt, V. R., Belrhali, H., & Sewell, B. T. (2010). Crystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocol. Protein expression and purification, 74(2), 211-216.en_ZA
dc.identifier.issn1046-5928en_ZA
dc.identifier.ris TY - Journal Article AU - van Rooyen, J M AU - Abratt, V R AU - Belrhali, H AU - Sewell, B T AB - Glutamine synthetase enzymes (GSs) are large oligomeric enzymes that play a critical role in nitrogen metabolism in all forms of life. To date, no crystal structures exist for the family of large (1 MDa) type III GS enzymes, which only share 9% sequence identity with the well characterized GSI and GSII enzymes. Here we present a novel protocol for the isolation of untagged Bacteroides fragilis GlnN expressed in an auxotrophic Escherichia coli strain. The rapid and scalable two-step protocol utilized differential precipitation by divalent cations followed by affinity chromatography to produce suitable quantities of homogenous material for structural characterization. Subsequent optimizations to the sample stability and solubility led to the discovery of conditions for the production of the first diffraction quality crystals of a type III GS enzyme. DA - 2010 DB - OpenUCT DP - University of Cape Town J1 - Protein Expression and Purification LK - https://open.uct.ac.za PB - University of Cape Town PY - 2010 SM - 1046-5928 T1 - Crystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocol TI - Crystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocol UR - http://hdl.handle.net/11427/20674 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/20674
dc.identifier.vancouvercitationvan Rooyen JM, Abratt VR, Belrhali H, Sewell BT. Crystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocol. Protein Expression and Purification. 2010; http://hdl.handle.net/11427/20674.en_ZA
dc.languageengen_ZA
dc.publisherElsevieren_ZA
dc.publisher.institutionUniversity of Cape Town
dc.rights.holderCreative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.sourceProtein Expression and Purificationen_ZA
dc.source.urihttp://www.journals.elsevier.com/protein-expression-and-purification
dc.subject.otherGlutamine synthetase type III GSIII Structure Bacteroides fragilis X-ray crystallography Protein pur
dc.titleCrystallization of recombinant Bacteroides fragilis glutamine synthetase (GlnN) isolated using a novel and rapid purification protocolen_ZA
dc.typeJournal Articleen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
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