Pipecolic acid is oxidized by renal and hepatic peroxisomes: Implications for Zellweger's cerebro-hepato-renal syndrome (CHRS)

doi:10.1016/0014-4827(86)90475-1

Experimental Cell Research

Volume 164, Issue 1, May 1986, Pages 267-271

https://doi.org/10.1016/0014-4827(86)90475-1 Get rights and content

Abstract

Increased levels of pipecolic acid have been reported in patients with cerebro-hepatorenal syndrom (CHRS) of Zellweger and the general deficiency of peroxisomal function has been implicated in its pathogenesis. We have therefore investigated the capacity of normal peroxisomes to metabolize pipecolic acid. Highly purified peroxisomes were obtained from rat liver and rat and beef kidney cortex by a recently developed method using metrizamide gradients and a vertical rotor. These preparations oxidized d,l-pipecolic acid as evidenced by the measurement of H₂O₂ production. Incubation with either the d- or l-isomer revealed that almost exclusively d-pipecolate is oxidized. The specific activities proved to be 20–50 times higher in renal than in hepatic peroxisomes. A commercially available crystalline suspension of d-amino acid oxidase from porcine kidney also oxidized the pipecolic acid with the following rates 54:36:1 respectively for d-:,d,l-:l-isomers. Incubation of vibratome sections of rat kidney and liver in a medium containing d-pipecolic acid and cerous ions, revealed electron-dense deposits over the matrix of peroxisomes confirming the localization also by fine structural cytochemistry.

These observations demonstrate the capability of mammalian peroxisomes to oxidize pipecolic acid and suggest that the absence or deficiency of peroxisomal d-amino acid oxidase may be implicated in the pathogenesis of hyperpipecolatemia in Zellweger's CHRS.

References (23)

J Dancis et al.
Comp biochem physiol
(1982)
DW Smith et al.
J pediatr
(1965)
E Passarge et al.
J ped
(1967)
YF Chang
Biochem biophys res commun
(1976)
YF Chang
J neurochem
(1978)
H Nishio et al.
Neurochem res
(1981)
JMF Trijbels et al.
J inher metab dis
(1979)
DW Arneson et al.
Arch neurol
(1982)
SL Goldfischer et al.
Science
(1973)
RI Kelley
Am j med genetics
(1983)

A Völkl et al.

Eur j biochem

(1985)

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Investigations on the occurrence and biochemical roles of free D‐amino acids and D‐amino acid‐containing peptides and proteins in living systems have increased in frequency and significance. Their occurrence and roles may vary substantially with progression from microbiotic to evermore advanced macrobiotic systems. We now understand many of the biosynthetic and regulatory pathways, which are outlined herein. Important uses for D‐amino acids in plants, invertebrates, and vertebrates are reviewed. Given its importance, a separate section on the occurrence and role of D‐amino acids in human disease is presented.
Mouse lysine catabolism to aminoadipate occurs primarily through the saccharopine pathway; implications for pyridoxine dependent epilepsy (PDE)
2017, Biochimica et Biophysica Acta - Molecular Basis of Disease
Citation Excerpt :
In point of fact, the 15N-AAA APE (%) in the plasma of mice injected with [ε-15N] lysine was not statistically different from the natural 15N-AAA levels found in mice injected with unlabelled lysine. This uncertainty about the functionality (or even existence) of the downstream steps of the pipecolate pathway is in keeping with recent similar experiment of lysine catabolic pathways [15] as well as with previous reports on low liver and kidney PIPOX activity [15,25,26]; it may be that pipecolic acid acts as a substrate in other cellular pathways yet to be delineated. Indeed, ε and α-15N-glutamic acid were not detected in blood samples but were found in similar levels in liver and brain at ~ 6 and 5 APE (%), respectively (data not shown).
Lysine is catabolized in mammals through the saccharopine and pipecolate pathways — the former is mainly hepatic and renal, and the latter is believed to play a role in the cerebral lysine oxidation. Both pathways lead to the formation of aminoadipic semialdehyde (AASA) that is then oxidized to aminoadipate (AAA) by antiquitin (ALDH7A1). Mutations in the ALDH7A1 gene result in the accumulation of AASA and its cyclic form, piperideine-6-carboxylate (P6C), which causes pyridoxine-dependent epilepsy (PDE). P6C reacts with pyridoxal 5′-phosphate (PLP) causing its inactivation. Here, we used liquid chromatography–mass spectrometry to investigate lysine catabolism in mice injected with lysine labelled at either its nitrogen epsilon (ε-¹⁵N) or nitrogen alpha (α-¹⁵N). Analysis of ε-¹⁵N and α-¹⁵N lysine catabolites in plasma, liver and brain suggested the saccharopine as the main pathway for AAA biosynthesis. Although there was evidence for upstream cerebral pipecolate pathway activity, the resulting pipecolate does not appear to be further oxidized into AASA/P6C/AAA. By far the bulk of lysine degradation and therefore, the primary source of lysine catabolites are hepatic and renal. The results indicate that the saccharopine pathway is primarily responsible for body's production of AASA/P6C. The centrality of the saccharopine pathway in whole body lysine catabolism opens new possibilities of therapeutic targets for PDE. We suggest that inhibition of this pathway upstream of AASA/P6C synthesis may be used to prevent its accumulation benefiting PDE patients. Inhibition of the enzyme aminoadipic semialdehyde synthase, for example, could constitute a new strategy to treat PDE and other inherited diseases of lysine catabolism.
Propiverine-induced accumulation of nuclear and cytosolic protein in F344 rat kidneys: Isolation and identification of the accumulating protein
2008, Toxicology and Applied Pharmacology
Male and female F344 rats but not B6C3F1 mice exposed for 104 weeks to propiverine hydrochloride (1-methylpiperid-4-yl 2,2-diphenyl-2-(1-propoxy)acetate hydrochloride), used for treatment of patients with neurogenic detrusor overactivity (NDO) and overactive bladder (OAB), presented with an accumulation of proteins in the cytosol and nuclei of renal proximal tubule epithelial cells, yet despite this, no increased renal tumor incidence was observed. In order to provide an improved interpretation of these findings and a better basis for human health risk assessment, male and female F344 rats were exposed for 16 weeks to 1000 ppm propiverine in the diet, the accumulating protein was isolated from the kidneys via cytosolic and nuclear preparations or laser-capture microdissection and analyzed using molecular weight determination and mass spectrometry. The accumulating protein was found to be d-amino acid oxidase (DAAO), an enzyme involved in amino and fatty acid metabolism. Subsequent reanalysis of kidney homogenate and nuclear samples as well as tissue sections using western blot and DAAO-immunohistochemistry, confirmed the presence and localization of DAAO in propiverine-treated male and female F344 rats. The accumulation of DAAO only in rats, and the limited similarity of rat DAAO with other species, including humans, suggests a rat-specific mechanism underlying the drug-induced renal DAAO accumulation with little relevance for patients chronically treated with propiverine.
Peroxisomes and oxidative stress
2006, Biochimica et Biophysica Acta - Molecular Cell Research
The discovery of the colocalization of catalase with H₂O₂-generating oxidases in peroxisomes was the first indication of their involvement in the metabolism of oxygen metabolites. In past decades it has been revealed that peroxisomes participate not only in the generation of reactive oxygen species (ROS) with grave consequences for cell fate such as malignant degeneration but also in cell rescue from the damaging effects of such radicals. In this review the role of peroxisomes in a variety of physiological and pathological processes involving ROS mainly in animal cells is presented. At the outset the enzymes generating and scavenging H₂O₂ and other oxygen metabolites are reviewed. The exposure of cultured cells to UV light and different oxidizing agents induces peroxisome proliferation with formation of tubular peroxisomes and apparent upregulation of PEX genes. Significant reduction of peroxisomal volume density and several of their enzymes is observed in inflammatory processes such as infections, ischemia–reperfusion injury and hepatic allograft rejection. The latter response is related to the suppressive effects of TNFα on peroxisomal function and on PPARα. Their massive proliferation induced by a variety of xenobiotics and the subsequent tumor formation in rodents is evidently due to an imbalance in the formation and scavenging of ROS, and is mediated by PPARα. In PEX5^−/− mice with the absence of functional peroxisomes severe abnormalities of mitochondria in different organs are observed which resemble closely those in respiratory chain disorders associated with oxidative stress. Interestingly, no evidence of oxidative damage to proteins or lipids, nor of increased peroxide production has been found in that mouse model. In this respect the role of PPARα, which is highly activated in those mice, in prevention of oxidative stress deserves further investigation.
l-Pipecolic acid oxidation in rat: subcellular localization and developmental study
1993, Biochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
By using a sensitive radioactive assay method, we present here evidence that l-pipecolic acid oxidase is localized in both mitochondria and peroxisomes of rat liver. Brain white matter contained a more than 2-fold higher activity of l-pipecolic acid oxidation than the brain cortex. Suborganellar fractionation studies indicate that while the enzyme is a matrix protein in mitochondria, it is membrane-associated in peroxisomes. Both rotenone and antimycin A completely inhibited the enzyme activity in mitochondria but not in peroxisomes. The enzyme was shown to be inducible in mitochondria and peroxisomes of rat liver and brain tissues by glucagon and di-(2-ethylhexyl)phthalate, respectively. We report here for the first time the developmental aspects of l-pipecolic acid oxidation activity in rat liver and brain tissues. l-Pipecolic acid oxidase activity was detectable in whole rat embryo at 10 days of gestation, suggesting active l-pipecolic acid metabolism early during development. In both liver and brain tissues l-pipecolic acid oxidation activity was highest at 15 days of gestation and decreased with age in prenatal and postnatal conditions.
Assay for l-pipecolate oxidase activity in human liver: detection of enzyme deficiency in hyperpipecolic acidaemia
1992, BBA - Molecular Basis of Disease
A direct assay method is described for l-pipecolate oxidase. The assay uses NaHSO₃ to trap the $L -α-amino[^{3} H]adipate$ δ-semialdehyde (AAS) formed as a direct reaction product of l-pipecolate oxidase from l-[³H]pipecolic acid. The adduct so formed was separated from the substrate on Dowex 50 (H⁺) column. The product was identified as [³H]AAS by amino acid analysis after breaking down the adduct by boiling under acidic conditions. The assay is simpler and more specific than fluorometric methods; it is also more sensitive; requiring at most 16 μg of liver peroxisome-enriched protein per assay. We have used this assay procedure to detect l-pipecolate oxidase in skin fibroblasts obtained from a control subject and from patients of hyperpipecolic acidaemia and Zellweger syndrome and found that this enzyme activity is present in the control, but absent or decreased in the patients with the peroxisomal disorders.

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An abstract of this report is presented at the 25th annual meeting of the American Society of Cell Biology in Atlanta, Georgia, USA, November 1985.

This study was supported by a grant (Fa 146/1–2) from the Deutsche Forschungsgemeinschaft, Bonn-Bad Godesberg, FRG.

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Short notePipecolic acid is oxidized by renal and hepatic peroxisomes: Implications for Zellweger's cerebro-hepato-renal syndrome (CHRS)☆

Abstract

Comp biochem physiol

J pediatr

J ped

Biochem biophys res commun

J neurochem

Neurochem res

J inher metab dis

Arch neurol

Science

Am j med genetics

Eur j biochem

Short note
Pipecolic acid is oxidized by renal and hepatic peroxisomes: Implications for Zellweger's cerebro-hepato-renal syndrome (CHRS)☆