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Functional studies of single-nucleotide polymorphic variants of human glutathione transferase T1-1 involving residues in the dimer interface
Stockholm University, Faculty of Science, Department of Neurochemistry.ORCID iD: 0000-0002-6416-064X
2011 (English)In: Archives of Biochemistry and Biophysics, ISSN 0003-9861, E-ISSN 1096-0384, Vol. 513, no 2, p. 87-93Article in journal (Refereed) Published
Abstract [en]

Glutathione transferase T1-1 catalyses detoxication and bioactivation processes in which glutathione conjugates are formed from endogenous and xenobiotic substrates, including alkylating agents and halogenated alkanes. Although the common null polymorphism of the human GSTT1 gene has been studied extensively, little is known about the consequences of GSTT1 single-nucleotide polymorphisms (SNPs). Here, we have examined the effects of two SNPs that alter amino acid residues in the dimer interface of the GST T1-1 protein and one that causes a conservative substitution in the core of the subunit. Variant proteins were expressed in an Escherichia coli strain in which the metabolism of ethylene dibromide to a glutathione conjugate leads to lacZ reversion mutations. We measured the kinetic properties of the enzymes with the characteristic substrate 1,2-epoxy-3-(p-nitrophenoxy)propane (EPNP) and determined the specific activities with several other substrates. Circular dichroism spectroscopy was used to measure protein thermal denaturation profiles. Variant T104P, which has been reported as inactive, showed weak but detectable activity with each substrate. Variant R76S was expressed at lower levels and showed much-reduced thermal stability. The results are interpreted in the context of the three-dimensional structure of human GST T1-1.

Place, publisher, year, edition, pages
2011. Vol. 513, no 2, p. 87-93
Keywords [en]
Glutathione transferase T1-1, Single nucleotide polymorphism, Ethylene dibromide
National Category
Natural Sciences
Identifiers
URN: urn:nbn:se:su:diva-66526DOI: 10.1016/j.abb.2011.07.003ISI: 000294376000003OAI: oai:DiVA.org:su-66526DiVA, id: diva2:469834
Note
authorCount :4Available from: 2011-12-27 Created: 2011-12-20 Last updated: 2022-02-24Bibliographically approved

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Mannervik, Bengt

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