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PMID: 1883325 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

A novel glutathione transferase (13-13) isolated from the matrix of rat liver mitochondria having structural similarity to class theta enzymes.

The Biochemical journal ·Vol. 278 ( Pt 1) ·1991-08-15 ·Pages 137-41

Harris JM, Meyer DJ, Coles B, Ketterer B

Abstract

A rat liver mitochondrial-matrix fraction was prepared and shown to have 1-chloro-2,4-dinitrobenzene(CDNB)-metabolizing glutathione transferase (GST) activity. Further fractionation by sequential gel filtration, isoelectric focusing or chromatofocusing and hydroxyapatite chromatography yielded three GSTs of pI 9.3, 8.9 and 7.5, none of which bound to a GSH-agarose affinity matrix. Most of the activity was associated with the pI-9.3 form, which was selected for further study. Its activity was tested with the following potential substrates in addition to CDNB: 1,2-dichloro-4-nitrobenzene, p-nitrobenzyl chloride, trans-4-phenylbut-3-en-2-one, 1,2-epoxy-3-(p-nitrophenoxy)propane, ethacrynic acid, menaphthyl sulphate, cumene hydroperoxide, linoleic acid hydroperoxide and 4-hydroxynon-2-enal. Appreciable activity was obtained only with CDNB and ethacrynic acid (82 and 26 mumol/min per mg of protein respectively). The apparent Km for GSH, using 1 mM-CDNB, was 1.9 mM. The enzyme is a dimer of subunit Mr 26,500. It has a free N-terminus, which has enabled the first 33 amino acids to be sequenced. This portion of primary structure has a sequence in common with members of the Theta class of GSTs (eg. 36% identity with subunit 12) and also a sequence which might function as a mitochondrial import signal. It is novel and has been named 'GST 13-13'.

MeSH Terms
Amino Acid Sequence Animals Chromatography Dinitrochlorobenzene/metabolism Glutathione Transferase/chemistry,isolation & purification Isoelectric Focusing Isoelectric Point Macromolecular Substances Male Microscopy, Electron Mitochondria, Liver/enzymology,ultrastructure Molecular Sequence Data Rats Rats, Inbred Strains Sequence Homology, Nucleic Acid
Chemicals
Dinitrochlorobenzene Macromolecular Substances Glutathione Transferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Harris J M
Department of Biochemistry, University College, London, U.K.
Meyer D J
Coles B
Ketterer B
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2780137
Published
1991-08-15
Pages
137-41
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1151459
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2780137 PMC Full Text View journal details
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