Rat glutathione S-transferase M4-4

An isoenzyme with unique structural features including a redox-reactive cysteine-115 residue that forms mixed disulphides with glutathione

H. Cheng, Tatyana L. Tchaikovskaya, Y. S L Tu, J. Chapman, B. Qian, W. M. Ching, M. Tien, J. D. Rowe, Y. V. Patskovsky, I. Listowsky, C. P D Tu

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Although the existence of the rat glutathione S-transferase (GST) M4 (rGSTM4) gene has been known for some time, the corresponding protein has not as yet been purified from tissue. A recombinant rGSTM4-4 was thus expressed in Escherichia coli from a chemically synthesized rGSTM4 gene. The catalytic efficiency (kcat/Km) of rGSTM4-4 for the 1-chloro-2,4-dinitrobenzene (CDNB) conjugation reaction was 50-180-fold less than that of the well-characterized homologous rGSTM1-1, and the pH optimum for the same reaction was 8.5 for rGSTM4-4 as opposed to 6.5 for rGSTM1-1. Molecular-modelling studies predict that key substitutions in the helix α4 region of rGSTM4-4 account for this pKa difference. A notable structural feature of rGSTM4-4 is the Cys- 115 residue in place of the Tyr-115 of other Mu-class GSTs. The thiol group of Cys- 115 is redox-reactive and readily forms a mixed disulphide even with GSH; the S-glutathiolated form of the enzyme is catalytically active. A mutated rGSTM4-4 (C115Y) had 6-10-fold greater catalytic efficiency than the wild-type rGSTM4-4. Trp-45, a conserved residue among Mu-class GSTs, is essential in rGSTM4-4 for both enzyme activity and binding to glutathione affinity matrices. Antibodies directed against either the unique C-terminal undecapeptide or tridecapeptide of rGSTM4 reacted with rat and mouse liver GSTs to reveal an orthologous mouse GSTM4-4 present at low basal levels but which is inducible in mouse liver. This subclass of rodent Mu GSTs with redox-active Cys-115 residues could have specialized physiological functions in response to oxidative stress.

Original languageEnglish (US)
Pages (from-to)403-414
Number of pages12
JournalBiochemical Journal
Volume356
Issue number2
DOIs
StatePublished - Jun 1 2001
Externally publishedYes

Fingerprint

Glutathione Disulfide
Glutathione Transferase
Disulfides
Isoenzymes
Oxidation-Reduction
Glutathione
Cysteine
Rats
Liver
Genes
Dinitrochlorobenzene
Oxidative stress
Molecular modeling
Enzyme activity
Enzymes
Sulfhydryl Compounds
Escherichia coli
Rodentia
Oxidative Stress
Substitution reactions

Keywords

  • Glutathiolation
  • Mass spectrometry
  • Oxidative stress
  • Structural model
  • Synthetic gene

ASJC Scopus subject areas

  • Biochemistry

Cite this

Rat glutathione S-transferase M4-4 : An isoenzyme with unique structural features including a redox-reactive cysteine-115 residue that forms mixed disulphides with glutathione. / Cheng, H.; Tchaikovskaya, Tatyana L.; Tu, Y. S L; Chapman, J.; Qian, B.; Ching, W. M.; Tien, M.; Rowe, J. D.; Patskovsky, Y. V.; Listowsky, I.; Tu, C. P D.

In: Biochemical Journal, Vol. 356, No. 2, 01.06.2001, p. 403-414.

Research output: Contribution to journalArticle

Cheng, H. ; Tchaikovskaya, Tatyana L. ; Tu, Y. S L ; Chapman, J. ; Qian, B. ; Ching, W. M. ; Tien, M. ; Rowe, J. D. ; Patskovsky, Y. V. ; Listowsky, I. ; Tu, C. P D. / Rat glutathione S-transferase M4-4 : An isoenzyme with unique structural features including a redox-reactive cysteine-115 residue that forms mixed disulphides with glutathione. In: Biochemical Journal. 2001 ; Vol. 356, No. 2. pp. 403-414.
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AU - Tchaikovskaya, Tatyana L.

AU - Tu, Y. S L

AU - Chapman, J.

AU - Qian, B.

AU - Ching, W. M.

AU - Tien, M.

AU - Rowe, J. D.

AU - Patskovsky, Y. V.

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