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Single-nucleotide p...
Single-nucleotide polymorphic variants of human glutathione transferase T1-1 differ in stability and functional properties
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Josephy, David (author)
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Kent, Meredith (author)
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- Mannervik, Bengt (author)
- Uppsala universitet,Institutionen för biokemi och organisk kemi
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(creator_code:org_t)
- Elsevier BV, 2009
- 2009
- English.
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In: Archives of Biochemistry and Biophysics. - : Elsevier BV. - 0003-9861 .- 1096-0384. ; 490:1, s. 24-29
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
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- We have previously expressed hexa-histidine-tagged human glutathione transferase GST T1-1 at very high levels in an Escherichia coli lacZ mutagenicity assay strain. Ethylene dibromide (EDB), which is activated by GST T1-1, produces a potent response in the mutation assay. We have now constructed and expressed two SNP variants of wild-type GST T1-1:D141N and E173K. The EDB activation activities of both variant enzymes, as measured by the lacZ mutagenicity assay, are greatly reduced The D141N variant behaved similarly to the wild-type enzyme, in terms of expression level and specific activities for conjugation of glutathione with 1,2-epoxy-3-(p-nitrophenoxy)propane (EPNP), ethylene diiodide (EDI), and 4-nitrobenzyl chloride (NBCl), and for peroxidative detoxication of cumene hydroperoxide (CuOOH). In contrast, variant E173K is poorly expressed, has no detectable activity with EPNP, NBCl, or CuOOH, and has EDI activity much lower than that of the wild-type enzyme. The circular dichroism (CD) thermal denaturation profiles of the wild-type protein and variant D141N show a sharp two-state transition between native and denatured states. Variant E173K showed a very different profile, consistent with improper or incomplete protein folding. Our results show that SNP variants can give rise to GSTT1-1 proteins with significantly altered properties. (C) 2009 Elsevier Inc. All rights reserved.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
Keyword
- Glutathione transferase
- Single nucleotide polymorphism
- Biology
- Biologi
Publication and Content Type
- ref (subject category)
- art (subject category)
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