The interorgan metabolism of glutathione

doi:10.1016/0020-711X(80)90005-1

International Journal of Biochemistry

Volume 12, Issue 4, 1980, Pages 545-551

https://doi.org/10.1016/0020-711X(80)90005-1 Get rights and content

Abstract

1.
1. Glutathione is the primary low molecular weight cellular thiol and a major reserve of cysteine. Glutathione is synthesized in the cytoplasm, where it participates in a variety of reactions as an electron donor or a nucleophilic cofactor.
2.
2. However, it now appears that the initial step in glutathione turnover is its release from the cell. The released glutathione is transported by the blood plasma to the kidney where it is efficiently extracted and degraded to its constituent amino acids.
3.
3. The initial reaction in the renal degradation of glutathione is catalyzed by y-glutamyltranspeptidase. which is localized on the external surface of the brush border membrane.
4.
4. Within the lumen of the renal proximal tubule, the γ-glutamyltranspeptidase apparently catalyzes a hydrolytic rather than a transpeptidation reaction.
5.
5. The resulting eysteinylglycine is hydrolyzed by two extracellular brush border membrane enzymes. aminopeptidase M and a recently identified peptidase.
6.
6. Reabsorption of the amino acids and redistribution of cysteine for protein synthesis or glutathione resynthesis completes the interorgan metabolism of glutathione.

References (47)

M.D. Armstrong
The occurrence of cystinylglycine in blood plasma
Biochim. biophys. Acta
(1979)
S. Bannai et al.
The export of glutathione from human diploid cells in culture
J. biol. Chem.
(1979)
R. Hahn et al.
The fate of extracellular glutathione in the rat
Biochim. biophys. Acta
(1978)
R.P. Hughey et al.
Comparison of the size and physical properties of γ-glutamyltranspeptidase purified from rat kidney following solubilization within papain or Triton X-100
J. biol. Chem.
(1976)
R.P. Hughey et al.
Specificity of a particulate rat renal peptidase and its localization along with other enzymes of mercapturic acid synthesis
Archs Biochem. Biophys.
(1978)
R.P. Hughey et al.
Comparison of the association and orientation of γ-glutamyltranspeptidase in lecithin vesicles and in native membranes
J. biol. Chem.
(1979)
D.P. Jones et al.
Metabolism of glutathione and a glutathione conjugate by isolated kidney cells
J. biol. Chem.
(1979)
A.M. Karkowsky et al.
Kinetic studies of sheep kidney γ-glutamyl transpeptidase
J. biol. Chem.
(1976)
T. Kuhlenschmidt et al.
Subcellular localization of rat kidney phosphate independent glutaminase
Archs Biochem. Biophys.
(1975)
T.M. McIntyre et al.
Comparison of the hydrolytic and transfer activities of rat renal γ-glutamyltranspeptidase
J. biol. Chem.
(1979)

A. Meister

Glutathione synthesis

S. Minato

Isolation of anthglutin, an inhibitor of γ-glutamyl transpeptidase, from penicillum oxalicum

Archs Biochem. Biophys.

(1979)

K. Ormstad et al.

Partial characterization of a glutathione oxidase present in rat kidney plasma membrane fraction

Biochem. biophys. Res. Commun.

(1979)

K. Ormstad et al.

Characteristics of glutathione biosynthesis by freshly isolated rat kidney cells

J. biol. Chem.

(1980)

P.G. Richman et al.

Regulation of γ-glutamyl-cysteine synthetase by nonallosteric feedback inhibition by glutathione

J. biol. Chem.

(1975)

J.D. Schulman et al.

Glutathionuria: Inborn error of metabolism due to tissue deficiency of gamma-glutamyi transpeptidase

Biochem. biophys. Res Commun.

(1975)

S.K. Srivastava et al.

The transport of oxidized glutathione from human erythrocytes

J. biol. Chem.

(1969)

S.S. Tate et al.

Interaction of γ-glutamyl transpeptidase with amino acids, dipeptides and derivatives and analogs of glutathione

J. biol. Chem.

(1974)

S.S. Tate et al.

Identity of maleate-stimulated glutaminase with γ-glutamyl transpeptidase in rat kidney

J. biol. Chem.

(1975)

S.S. Tate et al.

Conversion of glutathione to glutathione disulfide a catalytic function of γ-glutamyltranspeptidase

J. biol. Chem.

(1979)

G.A. Thompson et al.

Interrelationships between the binding site for amino acids, dipeptides and γ-glutamyl donors in γ-glutamyl transpeptidase

J. biol. Chem.

(1977)

H. Thor et al.

Effect of cysteine, N-acetylcysteine and methionine on glutathione biosynthesis and bromobenzene toxicity in isolated rat hepatocytes

Archs Biochem. Biophys.

(1979)

F. Tietze

Enzymatic method for quantitative determination of nanogram amounts of total and oxidized glutathione: application to mammalian blood and other tissues

Analyt. Biochem.

(1969)

Cited by (87)

Genetic mapping of renal glutathione suggests a novel regulatory locus on the murine X chromosome and overlap with hepatic glutathione regulation
2021, Free Radical Biology and Medicine
Citation Excerpt :
High GSH concentrations in the kidney are achieved via multiple mechanisms: normal intracellular synthesis and also import of circulating GSH [4], which mostly originates in the liver [5,6]. In fact, the kidneys are the primary consumer of circulating GSH [6,7], regularly hydrolyzing the GSH tripeptide to its constituent amino acids at the cell membrane, absorbing them, and then using those amino acids for GSH resynthesis or protein synthesis [8]. Robust levels of renal GSH are essential for preventing oxidative stress and progression of chronic renal diseases [9–12], as markers of ROS and oxidative damage are elevated in chronic kidney disease (CKD) [13,14].
Glutathione (GSH) is a critical cellular antioxidant that protects against byproducts of aerobic metabolism and other reactive electrophiles to prevent oxidative stress and cell death. Proper maintenance of its reduced form, GSH, in excess of its oxidized form, GSSG, prevents oxidative stress in the kidney and protects against the development of chronic kidney disease. Evidence has indicated that renal concentrations of GSH and GSSG, as well as their ratio GSH/GSSG, are moderately heritable, and past research has identified polymorphisms and candidate genes associated with these phenotypes in mice. Yet those discoveries were made with in silico mapping methods that are prone to false positives and power limitations, so the true loci and candidate genes that control renal glutathione remain unknown. The present study utilized high-resolution gene mapping with the Diversity Outbred mouse stock to identify causal loci underlying variation in renal GSH levels and redox status. Mapping output identified a suggestive locus associated with renal GSH on murine chromosome X at 51.602 Mbp, and bioinformatic analyses identified apoptosis-inducing factor mitochondria-associated 1 (Aifm1) as the most plausible candidate. Then, mapping outputs were compiled and compared against the genetic architecture of the hepatic GSH system, and we discovered a locus on murine chromosome 14 that overlaps between hepatic GSH concentrations and renal GSH redox potential. Overall, the results support our previously proposed model that the GSH redox system is regulated by both global and tissue-specific loci, vastly improving our understanding of GSH and its regulation and proposing new candidate genes for future mechanistic studies.
Oral administration of γ-glutamylcysteine increases intracellular glutathione levels above homeostasis in a randomised human trial pilot study
2017, Redox Biology
Citation Excerpt :
In this current human study, we investigated the potential systemic bioavailability of orally administered γ-GC. Dipeptides are not expected to be particularly useful as oral therapeutics as they are often readily hydrolysed by digestive or serum proteases; however, the unusual γ-glutamyl bond found in γ-GC is resistant to hydrolysis by most proteases and aminoproteases [34,35]. Animal safety trials have demonstrated γ-GC to be safe at limit acute and repeated doses [36].
To determine if orally dosed γ-glutamylcysteine (γ-GC) can increase cellular glutathione (GSH) levels above homeostasis. Many chronic and age-related disorders are associated with down-regulation, or impairment, of glutamate cysteine ligase (GCL). This suggests that γ-GC supply may become limiting for the maintenance of cellular GSH at the normal levels required to effectively protect against oxidative stress and any resulting physiological damage.
GSH levels were measured in lymphocytes of healthy, non-fasting participants before and after single oral doses (2 and 4 g) of γ-GC. Blood samples were immediately processed using high speed fluorescence-activated cell sorting to isolate 10⁶ lymphocytes that were then assayed for GSH content.
A single 2 g dose of γ-GC increased lymphocyte GSH content above basal levels (53±47%, p<0.01, n=14) within 90 min of administration. A randomized dosage (2 and 4 g γ-GC) crossover design was used to explore the pharmacokinetics of this GSH increase. In general, for both dose levels (n=9), GSH increased from initial basal levels over 3 h (t_max) before reaching maximum GSH concentrations (C_max) that were near two (2 g γ-GC) to three (4 g γ-GC) fold basal levels (0.4 nmol/10⁶ lymphocytes). Beyond t_max, GSH levels progressively declined reaching near basal levels by 5 h. The GSH half-life was between 2 and 3 h with exposure (AUC) to increased GSH levels of 0.7 (2 g γ-GC) and 1.8 (4 g γ-GC) nmol.h/10⁶ lymphocytes.
Oral γ-GC is a non-toxic form of cysteine that can be directly taken up by cells and transiently increase lymphocyte GSH above homeostatic levels. Our findings that γ-GC can increase GSH levels in healthy subjects suggests that it may have potential as an adjunct for treating diseases associated with chronic GSH depletion. This trial was registered at anzctr.org.au as ACTRN12612000952842.
Cytoprotective Systems within the Kidney
2010, Comprehensive Toxicology, Second Edition
The kidneys are particularly susceptible to injury from a diverse array of chemicals and drugs, including some that are therapeutic agents for which nephrotoxicity limits their efficacy. Additionally, several diseases and pathological states, such as diabetes and cardiovascular disease, exhibit renal dysfunction as a significant complication. Accordingly, identification of protective agents or development of strategies to prevent cell death or enhance cellular repair and/or proliferation is needed. This chapter will review cytoprotective mechanisms in renal proximal tubular cells, focusing on three classes of processes that have been the best studied: (1) thiol-disulfide redox protectants, including the glutathione and thioredoxin systems, metallothionein, N-acetyl-l-cysteine, and other low-molecular-weight thiols; (2) modulation of signaling pathways, involving epidermal growth factor, the mitogen-activated protein kinase family, protein kinase C and B (Akt), heat shock proteins, p53 and p21, peroxisome proliferator activator receptor-γ coactivator-1α (PGC-1α), BH3-family proteins, and the Nrf2 and NF-κB pathways; and (3) modulation of renal physiological function, focusing on intracellular pH and glycine and chloride channels. While this listing of cytoprotective mechanisms that can function in the kidneys is by no means complete, it demonstrates the diversity and breadth of protective systems that serve to balance the susceptibility of the kidneys, in particular the proximal tubular cells, and restore cellular and/or tissue function in disease states or after toxic insults.
Glutathione depletion and recovery after acute ethanol administration in the aging mouse
2007, Biochemical Pharmacology
Citation Excerpt :
The decreased levels of Cys observed in the kidney after 24 h in old mice may be indicative of a reduction in hepatic GSH biosynthesis and efflux in these animals. Renal Cys levels have been utilized as an indirect indicator of efflux since the majority of GSH transported from the sinusoidal membrane of the liver into the plasma ends up being degraded in the kidney by the enzyme γ-glutamyl transpeptidase resulting in the release of Cys [60]. Administration of radiolabelled GSH monoethylester resulted in quantitative increases in renal Cys concentration and in the labelling of the renal Cys pool to a similar specific activity as that of administered GSH [61].
Glutathione (GSH) plays an important role in the detoxification of ethanol (EtOH) and acute EtOH administration leads to GSH depletion in the liver and other tissues. Aging is also associated with a progressive decline in GSH levels and impairment in GSH biosynthesis in many tissues. Thus, the present study was designed to examine the effects of aging on EtOH-induced depletion and recovery of GSH in different tissues of the C57Bl/6NNIA mouse. EtOH (2–5 g/kg) or saline was administered i.p. to mice of ages 6 months (young), 12 months (mature), and 24 months (old); and GSH and cyst(e)ine concentrations were measured 0–24 h thereafter. EtOH administration (5 g/kg) depleted hepatic GSH levels >50% by 6 h in all animals. By 24 h, levels remained low in both young and old mice, but recovered to baseline levels in mature mice. At 6 h, the decrease in hepatic GSH was dose-dependent up to 3 g/kg EtOH, but not at higher doses. The extent of depletion at the 3 g/kg dose was dependent upon age, with old mice demonstrating significantly lower GSH levels than mature mice (P < 0.001). Altogether these results indicate that aging was associated with a greater degree of EtOH and fasting-induced GSH depletion and subsequent impaired recovery in liver. An impaired ability to recover was also observed in young animals. Further studies are required to determine if an inability to recover from GSH depletion by EtOH is associated with enhanced toxicity.
Role of glutathione transport processes in kidney function
2005, Toxicology and Applied Pharmacology
The kidneys are highly dependent on an adequate supply of glutathione (GSH) to maintain normal function. This is due, in part, to high rates of aerobic metabolism, particularly in the proximal tubules. Additionally, the kidneys are potentially exposed to high concentrations of oxidants and reactive electrophiles. Renal cellular concentrations of GSH are maintained by both intracellular synthesis and transport from outside the cell. Although function of specific carriers has not been definitively demonstrated, it is likely that multiple carriers are responsible for plasma membrane transport of GSH. Data suggest that the organic anion transporters OAT1 and OAT3 and the sodium–dicarboxylate 2 exchanger (SDCT2 or NaDC3) mediate uptake across the basolateral plasma membrane (BLM) and that the organic anion transporting polypeptide OATP1 and at least one of the multidrug resistance proteins mediate efflux across the brush-border plasma membrane (BBM). BLM transport may be used pharmacologically to provide renal proximal tubular cells with exogenous GSH to protect against oxidative stress whereas BBM transport functions physiologically in turnover of cellular GSH. The mitochondrial GSH pool is derived from cytoplasmic GSH by transport into the mitochondrial matrix and is mediated by the dicarboxylate and 2-oxoglutarate exchangers. Maintenance of the mitochondrial GSH pool is critical for cellular and mitochondrial redox homeostasis and is important in determining susceptibility to chemically induced apoptosis. Hence, membrane transport processes are critical to regulation of renal cellular and subcellular GSH pools and are determinants of susceptibility to cytotoxicity induced by oxidants and electrophiles.
Coincubation of tissue slices, a new way to study metabolic cooperation between organs: Hepatorenal cooperation in the biotransformation of CGP 47 969 A
2002, Journal of Pharmacological and Toxicological Methods
Introduction: There is limited information on in vitro/ex vivo tools to be used for studying interorgan metabolic cooperation. We report here the use of the tissue slice technique for this purpose. Methods: Rat liver and kidney slices were used to study metabolic cooperation for the metabolism of CGP 47 969, a potential anti-inflammatory compound which in vivo is extensively conjugated with glutathione and subsequently degraded via the mercapturic acid pathway. Results: Upon incubation with liver slices, CGP 47 969 was extensively conjugated with GSH whilst degradation of the GSH conjugate was moderate. Upon incubation with kidney slices, conjugation of CGP 47 969 with GSH was moderate but degradation of the GSH conjugate was complete. Upon coincubation of CGP 47 969 with liver and kidney slices, both conjugation with GSH and its subsequent degradation were almost complete. Thus, coincubation of liver and kidney slices permitted the efficient in vitro reproduction of the complete biotransformation of CGP 47 969 via its GSH conjugate to the ultimate mercapturic acid metabolite in a one step procedure. Discussion: This novel slice coincubation culture could serve as an in vitro model for interorgan cooperation in multistep metabolic processing.

View all citing articles on Scopus

View full text

MinireviewThe interorgan metabolism of glutathione

Abstract

Biochim. biophys. Acta

J. biol. Chem.

Biochim. biophys. Acta

J. biol. Chem.

Archs Biochem. Biophys.

J. biol. Chem.

J. biol. Chem.

J. biol. Chem.

Archs Biochem. Biophys.

J. biol. Chem.

Archs Biochem. Biophys.

Biochem. biophys. Res. Commun.

J. biol. Chem.

J. biol. Chem.

Biochem. biophys. Res Commun.

J. biol. Chem.

J. biol. Chem.

J. biol. Chem.

J. biol. Chem.

J. biol. Chem.

Archs Biochem. Biophys.

Analyt. Biochem.

Minireview
The interorgan metabolism of glutathione