RESUMO
Phospholipid oxidation generates several bioactive aldehydes that remain esterified to the glycerol backbone ('core' aldehydes). These aldehydes induce endothelial cells to produce monocyte chemotactic factors and enhance monocyte-endothelium adhesion. They also serve as ligands of scavenger receptors for the uptake of oxidized lipoproteins or apoptotic cells. The biochemical pathways involved in phospholipid aldehyde metabolism, however, remain largely unknown. In the present study, we have examined the efficacy of the three mammalian AKR (aldo-keto reductase) families in catalysing the reduction of phospholipid aldehydes. The model phospholipid aldehyde POVPC [1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphocholine] was efficiently reduced by members of the AKR1, but not by the AKR6 or the ARK7 family. In the AKR1 family, POVPC reductase activity was limited to AKR1A and B. No significant activity was observed with AKR1C enzymes. Among the active proteins, human AR (aldose reductase) (AKR1B1) showed the highest catalytic activity. The catalytic efficiency of human small intestinal AR (AKR1B10) was comparable with the murine AKR1B proteins 1B3 and 1B8. Among the murine proteins AKR1A4 and AKR1B7 showed appreciably lower catalytic activity as compared with 1B3 and 1B8. The human AKRs, 1B1 and 1B10, and the murine proteins, 1B3 and 1B8, also reduced C-7 and C-9 sn-2 aldehydes as well as POVPE [1-palmitoyl-2-(5-oxovaleroyl)-sn-glycero-3-phosphoethanolamine]. AKR1A4, B1, B7 and B8 catalysed the reduction of aldehydes generated in oxidized C(16:0-20:4) phosphatidylcholine with acyl, plasmenyl or alkyl linkage at the sn-1 position or C(16:0-20:4) phosphatidylglycerol or phosphatidic acid. AKR1B1 displayed the highest activity with phosphatidic acids; AKR1A4 was more efficient with long-chain aldehydes such as 5-hydroxy-8-oxo-6-octenoyl derivatives, whereas AKR1B8 preferred phosphatidylglycerol. These results suggest that proteins of the AKR1A and B families are efficient phospholipid aldehyde reductases, with non-overlapping substrate specificity, and may be involved in tissue-specific metabolism of endogenous or dietary phospholipid aldehydes.
Assuntos
Aldeído Redutase/metabolismo , Aldeídos/metabolismo , Isoenzimas/metabolismo , Fosfolipídeos/metabolismo , Aldeído Redutase/classificação , Aldeído Redutase/genética , Aldeídos/química , Animais , Humanos , Isoenzimas/genética , Camundongos , Dados de Sequência Molecular , Estrutura Molecular , Oxirredução , Fosfolipídeos/química , Especificidade por SubstratoRESUMO
The distribution of microbial aldo-keto reductases was examined and their immunochemical characterization was performed. p-Nitrobenzaldehyde, pyridine-3-aldehyde and ethyl 4-chloro-3-oxobutanoate reductase activities were found to be widely distributed in a variety of microorganisms. In immunodiffusion studies, most yeasts belonging to the genera Sporobolomyces, Sporidiobolus and Rhodotorula formed precipitin bands with anti-Sporobolomyces salmonicolor aldehyde reductase serum. Furthermore, the results of immunotitration experiments suggested that Sporobolomyces salmonicolor AKU 4429 contains other enzyme(s) which can reduce p-nitrobenzaldehyde, pyridine-3-aldehyde and/or ethyl 4-chloro-3-oxobutanoate, and which are inactivated by anti-Sporobolomyces salmonicolor aldehyde reductase serum.
Assuntos
Aldeído Redutase/análise , Leveduras/enzimologia , Aldeído Redutase/classificação , Aldeído Redutase/imunologia , Bactérias/enzimologia , Bactérias/imunologia , Reações Cruzadas , Fungos/enzimologia , Fungos/imunologia , Especificidade da Espécie , Leveduras/imunologiaRESUMO
Detoxication of the tobacco-specific carcinogen 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) in humans is mainly due to carbonyl reduction to the chiral alcohol 4-methylnitrosamino-1-(3-pyridyl)-1-butanol (NNAL), which undergoes glucuronidation and excretion. NNAL has a carcinogenic potential with (S)-NNAL being more tumorigenic in the mouse. Therefore, the enantioselectivity of NNK reductases seems toxicologically relevant. NNAL enantiomers were measured by a novel high-performance liquid chromatography procedure. The aldo-keto reductases AKR1C1, 1C2, and 1C4 and carbonyl reductase purified from human liver cytosol produced NNAL with >90% (S)-enantiomer in accordance with the enantioselectivity of NNK reduction by cytosol from liver, placenta, and lung. In contrast, the (R)-NNAL content was 35% on NNK reduction with 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) purified from human liver microsomes, but around 70% with human microsomes. The selectivity for (R)-NNAL formation was still higher with microsomes from placenta (87%) and lung (89% in 10 of 11 surgical samples). Microsomes from lung of one patient reduced NNK at a much lower rate, with production of 14% (R)-NNAL. This points to predominant reduction in microsomes by an enzyme with selectivity for (R)-NNAL formation that was apparently absent from the lung of one patient. Experiments with 18beta-glycyrrhetinic acid, a potent inhibitor of 11beta-HSD1, also indicated a minor or no role for 11beta-HSD1. Rat liver and lung microsomes produced NNAL with about 33% and 55% (R)-enantiomer and a mean contribution of 11beta-HSD1 of 12% and 32%, respectively. Multiple enzymes seem to participate in NNK reduction in human and rat tissues.