The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function

Simple item page
dc.contributor.authorAchilonu, Ikechukwu
dc.contributor.authorGildenhuys, Samantha
dc.contributor.authorFisher, Loren
dc.contributor.authorBurke, Jonathan
dc.contributor.authorFanucchi, Sylvia
dc.contributor.authorSewell, B Trevor
dc.contributor.authorFernandes, Manuel
dc.contributor.authorDirr, Heini W
dc.date.accessioned2016-01-26T06:10:43Z
dc.date.available2016-01-26T06:10:43Z
dc.date.issued2010
dc.date.updated2016-01-19T09:57:18Z
dc.description.abstractThe common fold shared by members of the glutathione-transferase (GST) family has a topologically conserved isoleucine residue at the N-terminus of helix 3 which is involved in the packing of helix 3 against two β-strands in domain 1. The role of the isoleucine residue in the structure, function and stability of GST was investigated by replacing the Ile71 residue in human GSTA1-1 by alanine or valine. The X-ray structures of the I71A and I71V mutants resolved at 1.75 and 2.51 Å, respectively, revealed that the mutations do not alter the overall structure of the protein compared with the wild type. Urea-induced equilibrium unfolding studies using circular dichroism and tryptophan fluorescence suggest that the mutation of Ile71 to alanine or valine reduces the stability of the protein. A functional assay with 1-chloro-2,4-dinitrobenzene shows that the mutation does not significantly alter the function of the protein relative to the wild type. Overall, the results suggest that conservation of the topologically conserved Ile71 maintains the structural stability of the protein but does not play a significant role in catalysis and substrate binding.en_ZA
dc.identifierhttp://dx.doi.org/10.1107/S1744309110019135
dc.identifier.apacitationAchilonu, I., Gildenhuys, S., Fisher, L., Burke, J., Fanucchi, S., Sewell, B. T., ... Dirr, H. W. (2010). The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function. <i>Acta Crystallographica Section F-Structural Biology and Crystallization Communications</i>, http://hdl.handle.net/11427/16541en_ZA
dc.identifier.chicagocitationAchilonu, Ikechukwu, Samantha Gildenhuys, Loren Fisher, Jonathan Burke, Sylvia Fanucchi, B Trevor Sewell, Manuel Fernandes, and Heini W Dirr "The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function." <i>Acta Crystallographica Section F-Structural Biology and Crystallization Communications</i> (2010) http://hdl.handle.net/11427/16541en_ZA
dc.identifier.citationAchilonu, I., Gildenhuys, S., Fisher, L., Burke, J., Fanucchi, S., Sewell, B., ... & Dirr, H. W. (2010). The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 66(7), 776-780.en_ZA
dc.identifier.issn1744-3091en_ZA
dc.identifier.ris TY - Journal Article AU - Achilonu, Ikechukwu AU - Gildenhuys, Samantha AU - Fisher, Loren AU - Burke, Jonathan AU - Fanucchi, Sylvia AU - Sewell, B Trevor AU - Fernandes, Manuel AU - Dirr, Heini W AB - The common fold shared by members of the glutathione-transferase (GST) family has a topologically conserved isoleucine residue at the N-terminus of helix 3 which is involved in the packing of helix 3 against two β-strands in domain 1. The role of the isoleucine residue in the structure, function and stability of GST was investigated by replacing the Ile71 residue in human GSTA1-1 by alanine or valine. The X-ray structures of the I71A and I71V mutants resolved at 1.75 and 2.51 Å, respectively, revealed that the mutations do not alter the overall structure of the protein compared with the wild type. Urea-induced equilibrium unfolding studies using circular dichroism and tryptophan fluorescence suggest that the mutation of Ile71 to alanine or valine reduces the stability of the protein. A functional assay with 1-chloro-2,4-dinitrobenzene shows that the mutation does not significantly alter the function of the protein relative to the wild type. Overall, the results suggest that conservation of the topologically conserved Ile71 maintains the structural stability of the protein but does not play a significant role in catalysis and substrate binding. DA - 2010 DB - OpenUCT DP - University of Cape Town J1 - Acta Crystallographica Section F-Structural Biology and Crystallization Communications LK - https://open.uct.ac.za PB - University of Cape Town PY - 2010 SM - 1744-3091 T1 - The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function TI - The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function UR - http://hdl.handle.net/11427/16541 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/16541
dc.identifier.vancouvercitationAchilonu I, Gildenhuys S, Fisher L, Burke J, Fanucchi S, Sewell BT, et al. The role of a topologically conserved isoleucine in glutathione transferase structure, stability and function. Acta Crystallographica Section F-Structural Biology and Crystallization Communications. 2010; http://hdl.handle.net/11427/16541.en_ZA
dc.languageengen_ZA
dc.publisherInternational Union of Crystallographyen_ZA
dc.publisher.departmentElectron Microscope Uniten_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.sourceActa Crystallographica Section F-Structural Biology and Crystallization Communicationsen_ZA
dc.source.urihttp://journals.iucr.org/f/
dc.subject.otherglutathione transferases
dc.titleThe role of a topologically conserved isoleucine in glutathione transferase structure, stability and functionen_ZA
dc.typeJournal Articleen_ZA
uct.subject.keywordsisoleucineen_ZA
uct.subject.keywordsglutathione transferase structureen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceArticleen_ZA
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Sewell_role_a_topologically_2010.pdf
Size:
654.55 KB
Format:
Adobe Portable Document Format
Description:
Download
License bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
license.txt
Size:
1.72 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Journal Articles