BACKGROUND: Cancer stem-like cells (CSCs) represent a subset of tumor cells that have the ability to self-renew, a long lifespan and a relatively quiescent phenotype, and show resistance to conventional therapies. Various markers are used to identify CSCs, and have shown that different CSC subtypes may be present within a tumor. One functional property of CSCs is their relative lack of proteasomal activity compared to the tumor bulk. METHODS: We introduced an unstable fluorescent molecule into FaDu oropharyngeal squamous cell carcinoma cells and analyzed the association of proteasome activity with aldehydehyde dehydrogenase (ALDH) activity as another common CSC marker, and with other stem-cell related properties of glucose metabolism. We also analyzed publicly available gene expression profiling data of ALDH+ CSCs for alterations in mRNAs associated with proteostasis. RESULTS: We show that FaDu CSCs identified by low proteasome activity are associated with the population identified by high ALDH activity. Futher characterization shows that these CSCs have a relatively high mitochondrial membrane potential and low levels of glucose transporter, indicating a non-Warburg metabolic phenotype. We also show that proteasome-low FaDu CSCs exhibit decreased rates of protein synthesis. Gene expression profiling of other cancer cell lines reveal common statistically significant differences in proteostasis in ALDH+ CSCs compared to the bulk of the tumor cells, including reduced levels of Hsp70 and/or Hsp90 in CSCs defined by ALDH, together with reduced levels of UCHL5 mRNA. CONCLUSIONS: These data provide additional insights into the functional characteristics of proteasome-low/ALDH-high CSCs, indicating a metabolic phenotype of reduced reliance on aerobic glycolysis and a decreased protein synthesis rate. We also identify specific chaperone and ubiquitin ligase activities that can be used to identify CSCs, with corresponding implications for therapeutic strategies that target CSCs through their altered metabolic properties.
- Klíčová slova
- Cancer stem cells, Glucose transporter, Mitochondrial membrane potential, Protein degradation, Proteosynthesis, Squamous cell carcinoma,
- MeSH
- aldehyddehydrogenasa * metabolismus genetika MeSH
- glukosa metabolismus MeSH
- lidé MeSH
- membránový potenciál mitochondrií MeSH
- mitochondrie * metabolismus MeSH
- nádorové buněčné linie MeSH
- nádorové kmenové buňky * metabolismus patologie MeSH
- proteasomový endopeptidasový komplex metabolismus MeSH
- proteiny usnadňující transport glukosy * metabolismus MeSH
- regulace genové exprese u nádorů MeSH
- spinocelulární karcinom * metabolismus patologie genetika MeSH
- stanovení celkové genové exprese MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- aldehyddehydrogenasa * MeSH
- glukosa MeSH
- proteasomový endopeptidasový komplex MeSH
- proteiny usnadňující transport glukosy * MeSH
ALDH7A1 deficiency is an epileptic encephalopathy whose seizures respond to treatment with supraphysiological doses of pyridoxine. It arises as a result of damaging variants in ALDH7A1, a gene in the lysine catabolism pathway. α-Aminoadipic semialdehyde (α-AASA) and Δ1-piperideine-6-carboxylate (P6C), which accumulate because of the block in the lysine pathway, are diagnostic biomarkers for this disorder. Recently, it has been reported that 6-oxo-pipecolic acid (6-oxo-PIP) also accumulates in the urine, CSF and plasma of ALDH7A1-deficient individuals and that, given its improved stability, it may be a more suitable biomarker for this disorder. This study measured 6-oxo-PIP in urine from a cohort of 30 patients where α-AASA was elevated and showed that it was above the normal range in all those above 6 months of age. However, 6-oxo-PIP levels were within the normal range in 33% of the patients below 6 months of age. Levels increased with age and correlated with a decrease in α-AASA levels. Longitudinal analysis of urine samples from ALDH7A1-deficient patients who were on a lysine restricted diet whilst receiving supraphysiological doses of pyridoxine showed that levels of 6-oxo-PIP remained elevated whilst α-AASA decreased. Similar to α-AASA, we found that elevated urinary excretion of 6-oxo-PIP can also occur in individuals with molybdenum cofactor deficiency. This study demonstrates that urinary 6-oxo-PIP may not be a suitable biomarker for ALDH7A1 deficiency in neonates. However, further studies are needed to understand the biochemistry leading to its accumulation and its potential long-term side effects.
- Klíčová slova
- 6‐oxo‐pipecolic acid, ALDH7A1 deficiency, aminoadipic semialdehyde, piperideine‐6‐carboxylate, pyridoxine‐dependent epilepsy,
- MeSH
- aldehyddehydrogenasa nedostatek genetika MeSH
- biologické markery * moč MeSH
- dítě MeSH
- epilepsie moč MeSH
- kojenec MeSH
- kyselina 2-aminoadipová moč analogy a deriváty MeSH
- kyseliny pipekolové * moč MeSH
- lidé MeSH
- lysin nedostatek moč MeSH
- mitochondriální aldehyddehydrogenasa nedostatek genetika MeSH
- novorozenec MeSH
- předškolní dítě MeSH
- pyridoxin nedostatek moč terapeutické užití MeSH
- Check Tag
- dítě MeSH
- kojenec MeSH
- lidé MeSH
- mužské pohlaví MeSH
- novorozenec MeSH
- předškolní dítě MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
- Názvy látek
- aldehyddehydrogenasa MeSH
- ALDH7A1 protein, human MeSH Prohlížeč
- biologické markery * MeSH
- kyselina 2-aminoadipová MeSH
- kyseliny pipekolové * MeSH
- lysin MeSH
- mitochondriální aldehyddehydrogenasa MeSH
- pipecolic acid MeSH Prohlížeč
- pyridoxin MeSH
Parrots produce stunning plumage colors through unique pigments called psittacofulvins. However, the mechanism underlying their ability to generate a spectrum of vibrant yellows, reds, and greens remains enigmatic. We uncover a unifying chemical basis for a wide range of parrot plumage colors, which result from the selective deposition of red aldehyde- and yellow carboxyl-containing psittacofulvin molecules in developing feathers. Through genetic mapping, biochemical assays, and single-cell genomics, we identified a critical player in this process, the aldehyde dehydrogenase ALDH3A2, which oxidizes aldehyde psittacofulvins into carboxyl forms in late-differentiating keratinocytes during feather development. The simplicity of the underlying molecular mechanism, in which a single enzyme influences the balance of red and yellow pigments, offers an explanation for the exceptional evolutionary lability of parrot coloration.
- MeSH
- aldehyddehydrogenasa * genetika MeSH
- barva MeSH
- biologické pigmenty * genetika metabolismus MeSH
- keratinocyty metabolismus MeSH
- oxidace-redukce MeSH
- papouškovití * anatomie a histologie genetika fyziologie MeSH
- peří * metabolismus MeSH
- pigmentace * genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- aldehyddehydrogenasa * MeSH
- biologické pigmenty * MeSH
- long-chain-aldehyde dehydrogenase MeSH Prohlížeč
Among the three active aldehyde oxidases in Arabidopsis thaliana leaves (AAO1-3), AAO3, which catalyzes the oxidation of abscisic-aldehyde to abscisic-acid, was shown recently to function as a reactive aldehyde detoxifier. Notably, aao2KO mutants exhibited less senescence symptoms and lower aldehyde accumulation, such as acrolein, benzaldehyde, and 4-hydroxyl-2-nonenal (HNE) than in wild-type leaves exposed to UV-C or Rose-Bengal. The effect of AAO2 expression absence on aldehyde detoxification by AAO3 and/or AAO1 was studied by comparing the response of wild-type plants to the response of single-functioning aao1 mutant (aao1S), aao2KO mutants, and single-functioning aao3 mutants (aao3Ss). Notably, aao3Ss exhibited similar aldehyde accumulation and chlorophyll content to aao2KO treated with UV-C or Rose-Bengal. In contrast, wild-type and aao1S exhibited higher aldehyde accumulation that resulted in lower remaining chlorophyll than in aao2KO leaves, indicating that the absence of active AAO2 enhanced AAO3 detoxification activity in aao2KO mutants. In support of this notion, employing abscisic-aldehyde as a specific substrate marker for AAO3 activity revealed enhanced AAO3 activity in aao2KO and aao3Ss leaves compared to wild-type treated with UV-C or Rose-Bengal. The similar abscisic-acid level accumulated in leaves of unstressed or stressed genotypes indicates that aldehyde detoxification by AAO3 is the cause for better stress resistance in aao2KO mutants. Employing the sulfuration process (known to activate aldehyde oxidases) in wild-type, aao2KO, and molybdenum-cofactor sulfurase (aba3-1) mutant plants revealed that the active AAO2 in WT employs sulfuration processes essential for AAO3 activity level, resulting in the lower AAO3 activity in WT than AAO3 activity in aao2KO.
- Klíčová slova
- Arabidopsis, Rose‐Bengal, UV‐C irradiation, aldehyde oxidase, aldehyde toxicity, reactive aldehydes, senescence, sulfuration,
- MeSH
- aldehydoxidasa metabolismus genetika MeSH
- aldehydy * metabolismus MeSH
- Arabidopsis * metabolismus genetika účinky záření MeSH
- chlorofyl metabolismus MeSH
- kyselina abscisová metabolismus MeSH
- listy rostlin * metabolismus genetika účinky záření MeSH
- mutace MeSH
- proteiny huseníčku * metabolismus genetika MeSH
- regulace genové exprese u rostlin MeSH
- ultrafialové záření * MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- AAO3 protein, Arabidopsis MeSH Prohlížeč
- aldehydoxidasa MeSH
- aldehydy * MeSH
- chlorofyl MeSH
- kyselina abscisová MeSH
- proteiny huseníčku * MeSH
Aldehyde dehydrogenases (ALDHs) represent a superfamily of enzymes, which oxidize aldehydes to the corresponding acids. Certain families, namely ALDH9 and ALDH10, are best active with ω-aminoaldehydes arising from the metabolism of polyamines such as 3-aminopropionaldehyde and 4-aminobutyraldehyde. Plant ALDH10s show broad specificity and accept many different aldehydes (aliphatic, aromatic and heterocyclic) as substrates. This work involved the above-mentioned aminoaldehydes acylated with dicarboxylic acids, phenylalanine, and tyrosine. The resulting products were then examined with native ALDH10 from pea and recombinant ALDH7s from pea and maize. This investigation aimed to find a common efficient substrate for the two plant ALDH families. One of the best natural substrates of ALDH7s is aminoadipic semialdehyde carrying a carboxylic group opposite the aldehyde group. The substrate properties of the new compounds were demonstrated by mass spectrometry of the reaction mixtures, spectrophotometric assays and molecular docking. The N-carboxyacyl derivatives were good substrates of pea ALDH10 but were only weakly oxidized by the two plant ALDH7s. The N-phenylalanyl and N-tyrosyl derivatives of 3-aminopropionaldehyde were good substrates of pea and maize ALDH7. Particularly the former compound was converted very efficiently (based on the kcat/Km ratio), but it was only weakly oxidized by pea ALDH10. Although no compound exhibited the same level of substrate properties for both ALDH families, we show that these enzymes may possess more common substrates than expected.
- Klíčová slova
- Acylation, Aldehyde dehydrogenase, Aminoaldehyde, Docking, Enzyme, Substrate,
- MeSH
- aldehyddehydrogenasa * metabolismus chemie genetika MeSH
- aldehydy * metabolismus chemie MeSH
- hrách setý * enzymologie MeSH
- kinetika MeSH
- kukuřice setá * enzymologie MeSH
- oxidace-redukce MeSH
- rostlinné proteiny metabolismus chemie genetika MeSH
- simulace molekulového dockingu * MeSH
- substrátová specifita MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- aldehyddehydrogenasa * MeSH
- aldehydy * MeSH
- rostlinné proteiny MeSH
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is known for its multifunctionality in several pathogenic bacteria. Our previously reported data suggest that the GAPDH homologue of Francisella tularensis, GapA, might also be involved in other processes beyond metabolism. In the present study, we explored GapA's potential implication in pathogenic processes at the host cell level. Using immunoelectron microscopy, we demonstrated the localization of this bacterial protein inside infected macrophages and its peripheral distribution in bacterial cells increasing with infection time. A quantitative proteomic approach based on stable isotope labeling of amino acids in cell culture (SILAC) combined with pull-down assay enabled the identification of several of GapA's potential interacting partners within the host cell proteome. Two of these partners were further confirmed by alternative methods. We also investigated the impact of gapA deletion on the transcription of selected cytokine genes and the activation of the main signaling pathways. Our results show that ∆gapA-induced transcription of genes encoding several cytokines whose expressions were not affected in cells infected with a fully virulent wild-type strain. That might be caused, at least in part, by the detected differences in ERK/MAPK signaling activation. The experimental observations together demonstrate that the F. tularensis GAPDH homologue is directly implicated in multiple host cellular processes and, thereby, that it participates in several molecular mechanisms of pathogenesis.
- Klíčová slova
- Francisella, glyceraldehyde-3-phosphate dehydrogenase, infection, interacting partners, multitasking, pleiotropy, secretion,
- MeSH
- cytokiny metabolismus MeSH
- exprese genu MeSH
- Francisella tularensis * genetika metabolismus MeSH
- glyceraldehyd-3-fosfátdehydrogenasy genetika metabolismus MeSH
- proteomika MeSH
- virulence genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cytokiny MeSH
- glyceraldehyd-3-fosfátdehydrogenasy MeSH
The Arabidopsis thaliana aldehyde oxidase 3 (AAO3) catalyzes the oxidation of abscisic aldehyde (ABal) to abscisic acid (ABA). Besides ABal, plants generate other aldehydes that can be toxic above a certain threshold. AAO3 knockout mutants (aao3) exhibited earlier senescence but equivalent relative water content compared with wild-type (WT) during normal growth or upon application of UV-C irradiation. Aldehyde profiling in leaves of 24-day-old plants revealed higher accumulation of acrolein, crotonaldehyde, 3Z-hexenal, hexanal and acetaldehyde in aao3 mutants compared with WT leaves. Similarly, higher levels of acrolein, benzaldehyde, crotonaldehyde, propionaldehyde, trans-2-hexenal and acetaldehyde were accumulated in aao3 mutants upon UV-C irradiation. Aldehydes application to plants hastened profuse senescence symptoms and higher accumulation of aldehydes, such as acrolein, benzaldehyde and 4-hydroxy-2-nonenal, in aao3 mutant leaves as compared with WT. The senescence symptoms included greater decrease in chlorophyll content and increase in transcript expression of the early senescence marker genes, Senescence-Related-Gene1, Stay-Green-Protein2 as well as NAC-LIKE, ACTIVATED-BY AP3/P1. Notably, although aao3 had lower ABA content than WT, members of the ABA-responding genes SnRKs were expressed at similar levels in aao3 and WT. Moreover, the other ABA-deficient mutants [aba2 and 9-cis-poxycarotenoid dioxygenase3-2 (nced3-2), that has functional AAO3] exhibited similar aldehydes accumulation and chlorophyll content like WT under normal growth conditions or UV-C irradiation. These results indicate that the absence of AAO3 oxidation activity and not the lower ABA and its associated function is responsible for the earlier senescence symptoms in aao3 mutant.
- Klíčová slova
- Arabidopsis, abscisic acid, aldehyde oxidase, reactive aldehydes, senescence,
- MeSH
- aldehydoxidasa genetika metabolismus MeSH
- aldehydy metabolismus toxicita MeSH
- Arabidopsis genetika fyziologie MeSH
- chlorofyl metabolismus MeSH
- kyselina abscisová metabolismus MeSH
- listy rostlin genetika fyziologie MeSH
- oxidace-redukce MeSH
- proteiny huseníčku genetika metabolismus MeSH
- regulátory růstu rostlin metabolismus MeSH
- senescence rostlin MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- AAO3 protein, Arabidopsis MeSH Prohlížeč
- aldehydoxidasa MeSH
- aldehydy MeSH
- chlorofyl MeSH
- kyselina abscisová MeSH
- proteiny huseníčku MeSH
- regulátory růstu rostlin MeSH
Temperature passively affects biological processes involved in plant growth. Therefore, it is challenging to study the dedicated temperature signalling pathways that orchestrate thermomorphogenesis, a suite of elongation growth-based adaptations that enhance leaf-cooling capacity. We screened a chemical library for compounds that restored hypocotyl elongation in the pif4-2-deficient mutant background at warm temperature conditions in Arabidopsis thaliana to identify modulators of thermomorphogenesis. The small aromatic compound 'Heatin', containing 1-iminomethyl-2-naphthol as a pharmacophore, was selected as an enhancer of elongation growth. We show that ARABIDOPSIS ALDEHYDE OXIDASES redundantly contribute to Heatin-mediated hypocotyl elongation. Following a chemical proteomics approach, the members of the NITRILASE1-subfamily of auxin biosynthesis enzymes were identified among the molecular targets of Heatin. Our data reveal that nitrilases are involved in promotion of hypocotyl elongation in response to high temperature and Heatin-mediated hypocotyl elongation requires the NITRILASE1-subfamily members, NIT1 and NIT2. Heatin inhibits NIT1-subfamily enzymatic activity in vitro and the application of Heatin accordingly results in the accumulation of NIT1-subfamily substrate indole-3-acetonitrile in vivo. However, levels of the NIT1-subfamily product, bioactive auxin (indole-3-acetic acid), were also significantly increased. It is likely that the stimulation of hypocotyl elongation by Heatin might be independent of its observed interaction with NITRILASE1-subfamily members. However, nitrilases may contribute to the Heatin response by stimulating indole-3-acetic acid biosynthesis in an indirect way. Heatin and its functional analogues present novel chemical entities for studying auxin biology.
- Klíčová slova
- 1-iminomethyl-2-naphthol, Arabidopsis, Heatin, IAN, NIT1-subfamily, PIF4, aldehyde oxidase, chemical genetics, indole-3-acetonitrile, nitrilases, thermomorphogenesis,
- MeSH
- aldehydoxidasa genetika metabolismus MeSH
- aminohydrolasy genetika metabolismus MeSH
- apomorfin analogy a deriváty farmakologie MeSH
- Arabidopsis účinky léků růst a vývoj MeSH
- herbicidy farmakologie MeSH
- hypokotyl účinky léků růst a vývoj MeSH
- inhibitory enzymů aplikace a dávkování chemie farmakologie MeSH
- kyseliny indoloctové MeSH
- molekulární struktura MeSH
- pikloram farmakologie MeSH
- proteiny huseníčku genetika metabolismus MeSH
- regulace genové exprese u rostlin účinky léků MeSH
- transkriptom účinky léků MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- 10,11-dihydroxy-N-n-propylnorapomorphine MeSH Prohlížeč
- AAO1 protein, Arabidopsis MeSH Prohlížeč
- aldehydoxidasa MeSH
- aminohydrolasy MeSH
- apomorfin MeSH
- herbicidy MeSH
- inhibitory enzymů MeSH
- kyseliny indoloctové MeSH
- nitrilase MeSH Prohlížeč
- pikloram MeSH
- proteiny huseníčku MeSH
S-nitrosation as a redox-based posttranslational modification of protein cysteine has emerged as an integral part of signaling pathways of nitric oxide across all types of organisms. Protein S-nitrosation status is controlled by two key mechanisms: by direct denitrosation performed by the thioredoxin/thioredoxin reductase system, and in an indirect way mediated by S-nitrosoglutathione reductase (GSNOR). GSNOR, which has been identified as a key component of S-nitrosothiols catabolism, catalyzes an irreversible decomposition of abundant intracellular S-nitrosothiol, S-nitrosoglutathione (GSNO) to oxidized glutathione using reduced NADH cofactor. In plants, GSNOR has been shown to play important roles in plant growth and development and plant responses to abiotic and biotic stress stimuli. In this chapter, optimized protocols of spectrophotometric measurement of GSNOR enzymatic activity and activity staining in native polyacrylamide gels in plant GSNOR are presented.
- Klíčová slova
- Nitric oxide, Plant stress, S-nitrosation, S-nitrosoglutathione reductase, S-nitrosothiols,
- MeSH
- aldehydoxidoreduktasy metabolismus MeSH
- barvení a značení metody MeSH
- enzymatické testy metody MeSH
- fluorescence MeSH
- NAD chemie MeSH
- nativní elektroforéza na polyakrylamidovém gelu MeSH
- nitrosace MeSH
- oxid dusnatý metabolismus MeSH
- průběh práce MeSH
- rostlinné extrakty izolace a purifikace metabolismus MeSH
- rostliny enzymologie MeSH
- S-nitrosoglutathion chemická syntéza chemie MeSH
- S-nitrosothioly metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- aldehydoxidoreduktasy MeSH
- formaldehyde dehydrogenase, glutathione-independent MeSH Prohlížeč
- NAD MeSH
- oxid dusnatý MeSH
- rostlinné extrakty MeSH
- S-nitrosoglutathion MeSH
- S-nitrosothioly MeSH
BACKGROUND: ALDH-2 has been considered an important molecular target for the treatment of drug addiction due to its involvement in the metabolism of the neurotransmitter dopamine: however, the molecular basis for the selective inhibition of ALDH-2 versus ALDH-1 should be better investigated to enable a more pragmatic approach to the design of novel ALDH-2 selective inhibitors. OBJECTIVE: In the present study, we investigated the molecular basis for the selective inhibition of ALDH-2 by the antioxidant isoflavonoid daidzin (IC50 = 0.15 μM) compared to isoform 1 of ALDH through molecular dynamics studies and semiempirical calculations of the enthalpy of interaction. METHODS: The applied methodology consisted of performing the molecular docking of daidzin in the structures of ALDH-1 and ALDH-2 and submitting the lower energy complexes obtained to semiempirical calculations and dynamic molecular simulations. RESULTS: Daidzin in complex with ALDH-2 presented directed and more specific interactions, resulting in stronger bonds in energetic terms and, therefore, in enthalpic gain. Moreover, the hydrophobic subunits of daidzin, in a conformationally more restricted environment (such as the catalytic site of ALDH-2), promote the better organization of the water molecules when immersed in the solvent, also resulting in an entropic gain. CONCLUSION: The molecular basis of selective inhibition of ALDH-2 by isoflavonoids and related compounds could be related to a more favorable equilibrium relationship between enthalpic and entropic features. The results described herein expand the available knowledge regarding the physiopathological and therapeutic mechanisms associated with drug addiction.
- Klíčová slova
- ALDH-2, daidzin, isoflavonoids, molecular dynamics, nucleus accumbens, selective inhibition,
- MeSH
- aldehyddehydrogenasa metabolismus MeSH
- dopamin metabolismus MeSH
- inhibitory enzymů farmakologie MeSH
- isoflavony farmakologie MeSH
- poruchy spojené s užíváním psychoaktivních látek farmakoterapie MeSH
- simulace molekulového dockingu MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- aldehyddehydrogenasa MeSH
- daidzin MeSH Prohlížeč
- dopamin MeSH
- inhibitory enzymů MeSH
- isoflavony MeSH
