Immunological Detection of Acetaldehyde-Protein Adducts in Ethanol-Treated Carrot Cells

Perata, Pierdomenico; Vernieri, Paolo; Armellini, D.; Bugnoli, M; Tognoni, F.; Alpi, Amedeo

doi:10.1104/pp.98.3.913

Cited by 36 publications

(20 citation statements)

References 26 publications

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“…It is well established that AA has two highly reactive pairs of electrons which have the potential to form covalent bonds to amino groups (Mauch et al, 1986). In other in vitro studies, AA formed adducts with various mammalian proteins such as tubulin, ribonuclease and BSA (Mauch et al, 1986;Jennett et al, 1987;Perata et al, 1992).…”

Section: Discussionmentioning

confidence: 99%

“…AA is a very reactive compound, capable of binding covalently to amino groups of proteins to form a Schiff base (Mauch et al, 1986;Perata et al, 1992). However, it seems that the AA molecule cannot easily penetrate the fruit epidermis; in tomato and grape, vapour penetration is mainly through the stem scar (Pesis and Frenkel, 1989;Pesis and Marinansky, 1993).…”

Section: Introductionmentioning

confidence: 99%

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Postharvest effects of acetaldehyde vapour on ripening-related enzyme activity in avocado fruit

Pesis

Faiman

Dori

1998

Postharvest Biology and Technology

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Exogenous application of acetaldehyde (AA) vapour to whole or half-peeled avocado fruits (Persea americana cv. 'Fuerte'), prior to storage, inhibited fruit ripening. This inhibition was characterized by a delay in fruit softening and a reduction in ethylene production. Activities of the cell wall depolymerizing enzymes polygalacturonase (PG), i-galactosidase (i-GAL) and endoglucanase (Cx), and of 1-aminocyclopropane-1-carboxylic acid oxidase (ACC oxidase), were reduced immediately after treatment. AA-treated fruit had reduced ethylene production and ACC oxidase activity, both in vivo and in vitro. Levels of total free sulfhydryl (SH) group compounds increased in AA-treated fruit, but diminished in untreated fruit. AA treatment inhibited fruit pulp oxidation, while untreated control fruits oxidized and became brown. Application of AA to a PG reaction mixture reduced PG activity on sodium polypectate. Pre-incubation of the substrate with AA did not result in reduced activity. Attempts to detect AA-bound proteins in AA-treated fruits were unsuccessful.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Postharvest effects of acetaldehyde vapour on ripening-related enzyme activity in avocado fruit

Pesis

Faiman

Dori

1998

Postharvest Biology and Technology

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“…Cytoplasmic PDC and ADH activities typically ferment pyruvate to ethanol in terrestrial plants (Dolferus et al, 1985;Mü cke et al, 1995;Kü rsteiner et al, 2003; for review, see Magneschi and Perata, 2009). PDC catalyzes the decarboxylation of pyruvate to acetaldehyde and CO 2 , while ADH reduces acetaldehyde to ethanol (Perata and Alpi, 1991;Perata et al, 1992), which is excreted from the organism. Chlamydomonas possesses three distinct enzymes that are potentially important for ethanol production when the cells become anoxic: ADH1 (a putative dualfunction alcohol/acetaldehyde dehydrogenase, previously annotated as ADHE because of its homology with the E. coli AdhE protein [Mus et al, 2007;Hemschemeier et al, 2008]; Joint Genome Institute [JGI] version 4.0 protein identifier 133318, Augustus version 5.0 protein identifier 518335) and two other putative alcohol dehydrogenases that we designate ADH2 (JGI version 4.0 protein identifier 121409, Augustus version 5.0 protein identifier 516421) and ADH3 (JGI version 4.0 protein identifier 82021, Augustus version 5.0 protein identifier 516422).…”

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confidence: 99%

A Mutant in theADH1Gene ofChlamydomonas reinhardtiiElicits Metabolic Restructuring during Anaerobiosis

et al. 2012

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The green alga Chlamydomonas reinhardtii has numerous genes encoding enzymes that function in fermentative pathways. Among these, the bifunctional alcohol/acetaldehyde dehydrogenase (ADH1), highly homologous to the Escherichia coli AdhE enzyme, is proposed to be a key component of fermentative metabolism. To investigate the physiological role of ADH1 in dark anoxic metabolism, a Chlamydomonas adh1 mutant was generated. We detected no ethanol synthesis in this mutant when it was placed under anoxia; the two other ADH homologs encoded on the Chlamydomonas genome do not appear to participate in ethanol production under our experimental conditions. Pyruvate formate lyase, acetate kinase, and hydrogenase protein levels were similar in wild-type cells and the adh1 mutant, while the mutant had significantly more pyruvate:ferredoxin oxidoreductase. Furthermore, a marked change in metabolite levels (in addition to ethanol) synthesized by the mutant under anoxic conditions was observed; formate levels were reduced, acetate levels were elevated, and the production of CO 2 was significantly reduced, but fermentative H 2 production was unchanged relative to wild-type cells. Of particular interest is the finding that the mutant accumulates high levels of extracellular glycerol, which requires NADH as a substrate for its synthesis. Lactate production is also increased slightly in the mutant relative to the control strain. These findings demonstrate a restructuring of fermentative metabolism in the adh1 mutant in a way that sustains the recycling (oxidation) of NADH and the survival of the mutant (similar to wild-type cell survival) during dark anoxic growth.

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“…The direct ELISA was tested for the assay of APA in a model system in which ovalbumin was used as a carrier protein for the synthesis of AOAs at acetaldehyde concen- trations from O to 1 mM (Israel et al, 1986;Perata et al, 1992). It was possible by the direct method to distinguish AOAl from other AOAs produced at lower acetaldehyde concentrations (Fig.…”

Section: Direct Elisa For the Assay Of Apamentioning

confidence: 99%

“…The production of acetaldehyde in tissue culture is thought to cause cell damage (Perata et al, 1988;Perata and Alpi, 1991). Perata et al (1992) tried to detect APA from suspension-cultured carrot cells using a direct ELISA method. However, APAs were detected only when both ethanol and a reducing agent (NaCNBH,) were supplied in the culture medium, and only trace amounts of endogenous APA were detected without the presence of a reducing agent.…”

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confidence: 99%

A Competitive Enzyme-Linked Immunosorbent Assay to Quantify Acetaldehyde-Protein Adducts That Accumulate in Dry Seeds during Aging

et al. 1997

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A competitive enzyme-linked immunosorbent assay (ELISA) was developed to quantify endogenous acetaldehyde-protein adducts (APAs) produced in plant seeds at low acetaldehyde concentrations without exogenous reducing agents. The key point of this technique is the use of a gelatin-acetaldehyde adduct, which is synthesized under 1 mM acetaldehyde and 10 mM NaCNBH,, to pre-coat plate wells to obtain the proper binding parameters for the quantification of APA in seed proteins. Compared with the traditional, direct ELISA method, the competitive one has higher sensitivity and less background. Using competitive ELISA, we determined the accumulation of endogenous APAs in seeds in relation to the loss of seed viability. Lettuce seeds were exposed to 2 mM gaseous acetaldehyde during storage for 30 or 45 d; the relative humidity and temperature of storage were studied independently. Viability decreased only in acetaldehyde-treated seeds, as either the temperature or the relative humidity increased. A loss in viability was accompanied by an increase in the accumulation of APA. The APA content also increased as viability decreased in five species of seeds, which were aged naturally without exposure to acetaldehyde. It is suggested that the modification of functional seed proteins with endogenously evolved acetaldehyde may be an important cause of seed aging.Acetaldehyde is an intermediate of alcoholic fermentation in plants. The toxic effects of acetaldehyde may arise from its nonenzymatic binding with proteins (Gaines et al., 1977;Stevens et al., 1981;Donohue et al., 1983) and DNA (Restow and Obe, 1978;Fraenkel-Conrat and Singer, 1988). Israel et al. (1986) found that the immunization of mice with acetaldehyde conjugated to human plasma proteins results in the production of a polyclonal antibody, which reacts with various APAs. The production of acetaldehyde in tissue culture is thought to cause cell damage (Perata et al., 1988;Perata and Alpi, 1991). Perata et al. (1992) tried to detect APA from suspension-cultured carrot cells using a direct ELISA method. However, APAs were detected only when both ethanol and a reducing agent (NaCNBH,) were supplied in the culture medium, and only trace amounts of endogenous APA were detected without the presence of a reducing agent. Among the many kinds of volatile compounds that have evolved from dry seeds, acetaldehyde and ethanol are common components in the various seed species (Zhang et al., 1993). Acetaldehyde is effective in causing the rapid deterioration of seeds when it is applied as a gas to seeds during storage periods (Zhang et al., 1994). We proposed a hypothesis that seed aging may be caused by a modification of seed proteins by endogenously produced acetaldehyde (Zhang et al., 1995). In this paper we report the detection of endogenous APA from five species of dry seeds using a sensitive, competitive ELISA technique and demonstrate the increased accumulation of APA in aged seeds. MATERIALS AND METHODS Seed Materials and Storage MethodsSeeds with high viability (90% or more) ...

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Immunological Detection of Acetaldehyde-Protein Adducts in Ethanol-Treated Carrot Cells

Cited by 36 publications

References 26 publications

Postharvest effects of acetaldehyde vapour on ripening-related enzyme activity in avocado fruit

Postharvest effects of acetaldehyde vapour on ripening-related enzyme activity in avocado fruit

A Mutant in theADH1Gene ofChlamydomonas reinhardtiiElicits Metabolic Restructuring during Anaerobiosis

A Competitive Enzyme-Linked Immunosorbent Assay to Quantify Acetaldehyde-Protein Adducts That Accumulate in Dry Seeds during Aging

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