Fish and mammalian liver cytosolic glutathione S-transferases: Substrate specificities and immunological comparison

The glutathione S-transferases that were purified to homogeneity from liver cytosol have overlapping but distinct substrate specificities and different isoelectric points. This report explores the possibility of using preparative electrofocusing to compare the composition of the transferases in liver and kidney cytosol. Hepatic cytosol from adult male Sprague–Dawley rats was resolved by isoelectric focusing on Sephadex columns into five peaks of transferase activity, each with characteristic substrate specificity. The first four peaks of transferase activity (in order of decreasing basicity) are identified as transferases AA, B, A and C respectively, on the basis of substrate specificity, but the fifth peak (pI6.6) does not correspond to a previously described transferase. Isoelectric focusing of renal cytosol resolves only three major peaks of transferase activity, each with narrow substrate specificity. In the kidney, peak 1 (pI9.0) has most of the activity toward 1-chloro-2,4-dinitrobenzene, peak 2 (pI8.5) toward p-nitrobenzyl chloride, and peak 3 (pI7.0) toward trans-4-phenylbut-3-en-2-one. Renal transferase peak 1 (pI9.0) appears to correspond to transferase B on the basis of pI, substrate specificity and antigenicity. Kidney transferase peaks 2 (pI8.5) and 3 (pI7.0) do not correspond to previously described glutathione S-transferases, although kidney transferase peak 3 is similar to the transferase peak 5 from focused hepatic cytosol. Transferases A and C were not found in kidney cytosol, and transferase AA was detected in only one out of six replicates. Thus it is important to recognize the contribution of individual transferases to total transferase activity in that each transferase may be regulated independently.

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Novel functional association of rat testicular membrane-associated cytosolic glutathione S transferases and cyclooxygenase in vitro

Asian Journal of Andrology ◽

10.1111/j.1745-7262.2005.00030.x ◽

2005 ◽

Vol 7 (2) ◽

pp. 171-178 ◽

Cited By ~ 2

Author(s):

S. Neeraja ◽

B. Ramakrishna ◽

A. S. Sreenath ◽

G. V. Reddy ◽

P. R. K. Reddy ◽

...

Keyword(s):

Functional Association ◽

Cytosolic Glutathione ◽

Glutathione S Transferases

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Human glutathione S-transferases. The Ha multigene family encodes products of different but overlapping substrate specificities.

Journal of Biological Chemistry ◽

10.1016/s0021-9258(18)37626-9 ◽

1988 ◽

Vol 263 (26) ◽

pp. 12797-12800

Author(s):

N W Chow ◽

J Whang-Peng ◽

C S Kao-Shan ◽

M F Tam ◽

H C Lai ◽

...

Keyword(s):

Multigene Family ◽

Substrate Specificities ◽

Glutathione S Transferases

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Glutathione S-transferases from the gastrointestinal nematode Heligmosomoides polygyrus and mammalian liver compared

Comparative Biochemistry and Physiology Part B Comparative Biochemistry ◽

10.1016/0305-0491(94)90121-x ◽

1994 ◽

Vol 109 (4) ◽

pp. 585-592 ◽

Cited By ~ 11

Author(s):

Peter M. Brophy ◽

Anne Ben-Smith ◽

Alan Brown ◽

Jerzy M. Behnke ◽

David I. Pritchard

Keyword(s):

Gastrointestinal Nematode ◽

Heligmosomoides Polygyrus ◽

Mammalian Liver ◽

Glutathione S Transferases

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Mass spectrometric analysis of rat liver cytosolic glutathione S-transferases: modifications are limited to N-terminal processing

Biochemical Journal ◽

10.1042/bj3080069 ◽

1995 ◽

Vol 308 (1) ◽

pp. 69-75 ◽

Cited By ~ 14

Author(s):

H I Yeh ◽

C H Hsieh ◽

L Y Wang ◽

S P Tsai ◽

H Y Hsu ◽

...

Keyword(s):

Molecular Mass ◽

Female Rats ◽

Spectrometric Analysis ◽

Affinity Column ◽

Female Rat ◽

Significant Difference ◽

Cytosolic Glutathione ◽

Glutathione S Transferases ◽

Molecular Masses ◽

Rat Livers

Cytosolic glutathione S-transferases (GSTs) from rat livers were purified using an S-hexylglutathione affinity column. The GST subunits were resolved by reverse-phase HPLC and their molecular masses were determined by electrospray mass spectrometry. The major hepatic GSTs detected were subunits 1, 1′, 2, 3 and 4, with molecular mass of 25,520, 25,473, 25,188, 25,782 and 25,571 Da respectively. Subunits 6, 7 and 10 are minor components, with molecular mass of 25,551, 23,308 and 25,211 Da respectively. Alternatively, the hepatic GSTs were purified using a glutathione affinity column. Subunits 1, 1′, 2, 8 and 10 were eluted from this column with GSSG, the oxidized form of glutathione. Subunit 8 has a molecular mass of 25,553 Da. The remaining proteins on the glutathione affinity column were removed with glutathione and S-hexylglutathione. Subunits 2, 3, 4 and 6 could be detected in the eluate. We could not detect any significant difference in molecular mass between GSTs isolated from male and female rat livers. Cytosolic GSTs were isolated from livers of buthionine sulphoximine-treated female rats for MS analysis. The molecular masses obtained were identical to those determined for the controls.

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Cytosolic glutathione S-transferases of Oesophagostomum dentatum

Parasitology ◽

10.1017/s0031182008004769 ◽

2008 ◽

Vol 135 (10) ◽

pp. 1215-1223 ◽

Cited By ~ 9

Author(s):

A. JOACHIM ◽

B. RUTTKOWSKI

Keyword(s):

Amino Acids ◽

Pcr Primers ◽

Mrna Levels ◽

Fourth Stage ◽

Glutathione S Transferase ◽

Cdna Sequences ◽

Oesophagostomum Dentatum ◽

Cytosolic Glutathione ◽

Glutathione S Transferases

SUMMARYOesophagostomum dentatum stages were investigated for glutathione S-transferase (GST) expression at the protein and mRNA levels. GST activity was detected in all stages (infectious and parasitic stages including third- and fourth-stage larvae of different ages as well as males and females) and could be dose-dependently inhibited with sulfobromophthalein (SBP). Addition of SBP to in vitro larval cultures reversibly inhibited development from third- to fourth-stage larvae. Two glutathione-affinity purified proteins (23 and 25 kDa) were detected in lysates of exsheathed third-stage larvae by SDS-PAGE. PCR-primers were designed based on peptide sequences and conserved GST sequences of other nematodes for complete cDNA sequences (621 and 624 nt) of 2 isoforms, Od-GST1 and Od-GST2, with 72% nucleotide similarity and 75% for the deduced proteins. Genomic sequences consisted of 7 exons and 6 introns spanning 1296 bp for Od-GST1 and 1579 and 1606 bp for Od-GST2. Quantitative real-time-PCR revealed considerably elevated levels of Od-GST1 in the early parasitic stages and slightly reduced levels of Od-GST2 in male worms. Both Od-GSTs were most similar to GST of Ancylostoma caninum (nucleotides: 73 and 70%; amino acids: 80 and 73%). The first three exons (75 amino acids) corresponded to a synthetic prostaglandin D2 synthase (53% similarity). O. dentatum GSTs might be involved in intrinsic metabolic pathways which could play a role both in nematode physiology and in host-parasite interactions.

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Purification of three cytosolic glutathione S-transferases from adult Schistosoma mansoni

Archives of Biochemistry and Biophysics ◽

10.1016/0003-9861(88)90563-2 ◽

1988 ◽

Vol 264 (1) ◽

pp. 1-12 ◽

Cited By ~ 32

Author(s):

Kathleen A. O'Leary ◽

James W. Tracy

Keyword(s):

Schistosoma Mansoni ◽

Cytosolic Glutathione ◽

Glutathione S Transferases

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Induction of cytosolic Glutathione S-transferases from Atlantic eel (Anguilla anguilla) after intraperitoneal treatment with polychlorinated biphenyls

The Science of The Total Environment ◽

10.1016/s0048-9697(02)00130-4 ◽

2002 ◽

Vol 297 (1-3) ◽

pp. 141-151 ◽

Cited By ~ 16

Author(s):

M Pérez López ◽

M.C Nóvoa Valiñas ◽

M.J Melgar Riol

Keyword(s):

Polychlorinated Biphenyls ◽

Anguilla Anguilla ◽

Cytosolic Glutathione ◽

Glutathione S Transferases ◽

Intraperitoneal Treatment

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Human intestinal glutathione S-transferases

Biochemical Journal ◽

10.1042/bj2570471 ◽

1989 ◽

Vol 257 (2) ◽

pp. 471-476 ◽

Cited By ~ 35

Author(s):

W H M Peters ◽

H M J Roelofs ◽

F M Nagengast ◽

J H M van Tongeren

Keyword(s):

Small Intestine ◽

Large Intestine ◽

Specific Activity ◽

Distal Colon ◽

Glutathione S Transferase ◽

Distal Small Intestine ◽

Cytosolic Glutathione ◽

Glutathione S Transferases ◽

Isoenzyme Composition ◽

Specific Activities

Cytosolic glutathione S-transferases were purified from the epithelial cells of human small and large intestine. These preparations were characterized with regard to specific activities, subunit and isoenzyme composition. Isoenzyme composition and specific activity showed little variation from proximal to distal small intestine. Specific activities of hepatic and intestinal enzymes from the same patient were comparable. Hepatic enzymes were mainly composed of 25 kDa subunits. Transferases from small intestine contained 24 and 25 kDa subunits, in variable amounts. Colon enzymes were composed of 24 kDa subunits. In most preparations, however, minor amounts of 27 and 27.5 kDa subunits were detectable. Separation into isoforms by isoelectric focusing revealed striking differences: glutathione S-transferases from liver were mainly basic or neutral, enzymes from small intestine were basic, neutral and acidic, whereas large intestine contained acidic isoforms only. The intestinal acidic transferase most probably was identical with glutathione S-transferase Pi, isolated from human placenta. In the hepatic preparation, this isoform was hardly detectable. The specific activity of glutathione S-transferase showed a sharp fall from small to large intestine. In proximal and distal colon, activity seemed to be about equal. In the ascending colon there might be a relationship between specific activity of glutathione S-transferases and age of the patient, activity decreasing with increasing age.

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