Polyamines participate in the processes of cell growth and development. The degradation branch of their metabolism involves amine oxidases. The oxidation of spermine, spermidine and putrescine releases hydrogen peroxide and the corresponding aminoaldehyde. Polyamine-derived aminoaldehydes have been found to be cytotoxic, and they represent the subject of this review. 3-aminopropanal disrupts the lysosomal membrane and triggers apoptosis or necrosis in the damaged cells. It is implicated in the pathogenesis of cerebral ischemia. Furthermore, 3-aminopropanal yields acrolein through the elimination of ammonia. This reactive aldehyde is also generated by the decomposition of aminoaldehydes produced in the reaction of serum amine oxidase with spermidine or spermine. In addition, acrolein is a common environmental pollutant. It causes covalent modifications of proteins, including carbonylation, the production of Michael-type adducts and cross-linking, and it has been associated with inflammation-related diseases. APAL and acrolein are detoxified by aldehyde dehydrogenases and other mechanisms. High-performance liquid chromatography, immunochemistry and mass spectrometry have been largely used to analyze the presence of polyamine-derived aminoaldehydes and protein modifications elicited by their effect. However, the main and still open challenge is to find clues for discovering clear linkages between aldehyde-induced modifications of specific proteins and the development of various diseases.

• Klíčová slova
• 3-aminopropanal, Michael adduct, Schiff base, acrolein, aldehyde dehydrogenase, amine oxidase, aminoaldehyde, cytotoxicity, glutathione, protein modification,
• MeSH
• akrolein * farmakologie   MeSH
• aldehydy farmakologie   MeSH
• polyaminy * MeSH
• spermidin farmakologie   MeSH
• spermin farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• 3-aminopropionaldehyde MeSH Prohlížeč
• akrolein * MeSH
• aldehydy MeSH
• polyaminy * MeSH
• spermidin MeSH
• spermin MeSH

The crystal structures of both isoforms of the aminoaldehyde dehydrogenase from pea (PsAMADH) have been solved recently [Tylichováet al. (2010) J Mol Biol396, 870-882]. The characterization of the PsAMADH2 proteins, altered here by site-directed mutagenesis, suggests that the D110 and D113 residues at the entrance to the substrate channel are required for high-affinity binding of ω-aminoaldehydes to PsAMADH2 and for enzyme activity, whereas N162, near catalytic C294, contributes mainly to the enzyme's catalytic rate. Inside the substrate cavity, W170 and Y163, and, to a certain extent, L166 and M167 probably preserve the optimal overall geometry of the substrate channel that allows for the appropriate orientation of the substrate. Unconserved W288 appears to affect the affinity of the enzyme for the substrate amino group through control of the substrate channel diameter without affecting the reaction rate. Therefore, W288 may be a key determinant of the differences in substrate specificity found among plant AMADH isoforms when they interact with naturally occurring substrates such as 3-aminopropionaldehyde and 4-aminobutyraldehyde.

• MeSH
• aldehydoxidoreduktasy chemie   genetika   izolace a purifikace   metabolismus   MeSH
• aldehydy metabolismus   MeSH
• aminokyseliny aromatické metabolismus   MeSH
• aminokyseliny dikarboxylové metabolismus   MeSH
• betain analogy a deriváty   metabolismus   MeSH
• biokatalýza MeSH
• cirkulární dichroismus MeSH
• hrách setý enzymologie   MeSH
• izoenzymy metabolismus   MeSH
• katalytická doména MeSH
• kinetika MeSH
• mutageneze cílená MeSH
• mutantní proteiny chemie   izolace a purifikace   metabolismus   MeSH
• propylaminy metabolismus   MeSH
• rekombinantní proteiny chemie   izolace a purifikace   metabolismus   MeSH
• rostlinné proteiny chemie   genetika   izolace a purifikace   metabolismus   MeSH
• sekundární struktura proteinů MeSH
• substrátová specifita MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 3-aminopropionaldehyde MeSH Prohlížeč
• 4-aminobutyraldehyde MeSH Prohlížeč
• aldehydoxidoreduktasy MeSH
• aldehydy MeSH
• aminokyseliny aromatické MeSH
• aminokyseliny dikarboxylové MeSH
• betain MeSH
• betaine aldehyde MeSH Prohlížeč
• izoenzymy MeSH
• mutantní proteiny MeSH
• propylaminy MeSH
• rekombinantní proteiny MeSH
• rostlinné proteiny MeSH

Plant aminoaldehyde dehydrogenases (AMADHs, EC 1.2.1.19) belong to the family 10 of aldehyde dehydrogenases and participate in the metabolism of compounds related to amino acids such as polyamines or osmoprotectants. Their broad specificity covers ω-aminoaldehydes, aliphatic and aromatic aldehydes as well as nitrogen-containing heterocyclic aldehydes. The substrate preference of plant AMADHs is determined by the presence of aspartic acid and aromatic residues in the substrate channel. In this work, 15 new N-acyl derivates of 3-aminopropanal (APAL) and 4-aminobutanal (ABAL) were synthesized and confirmed as substrates of two pea AMADH isoenzymes (PsAMADH 1 and 2). The compounds were designed considering the previously demonstrated conversion of N-acetyl derivatives as well as substrate channel dimensions (5-8 Å × 14 Å). The acyl chain length and its branching were found less significant for substrate properties than the length of the initial natural substrate. In general, APAL derivatives were found more efficient than the corresponding ABAL derivatives because of the prevailing higher conversion rates and lower K m values. Differences in enzymatic performance between the two isoenzymes corresponded in part to their preferences to APAL to ABAL. The higher PsAMADH2 affinity to substrates correlated with more frequent occurrence of an excess substrate inhibition. Molecular docking indicated the possible auxiliary role of Tyr163, Ser295 and Gln451 in binding of the new substrates. The only derivative carrying a free carboxyl group (N-adipoyl APAL) was surprisingly better substrate than ABAL in PsAMADH2 reaction indicating that also negatively charged aldehydes might be good substrates for ALDH10 family.

• MeSH
• aldehyddehydrogenasa chemie   metabolismus   MeSH
• aldehydy chemie   metabolismus   MeSH
• hrách setý chemie   enzymologie   MeSH
• kinetika MeSH
• molekulární struktura MeSH
• propylaminy chemie   metabolismus   MeSH
• rostlinné proteiny chemie   metabolismus   MeSH
• simulace molekulového dockingu MeSH
• substrátová specifita MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 3-aminopropionaldehyde MeSH Prohlížeč
• aldehyddehydrogenasa MeSH
• aldehydy MeSH
• propylaminy MeSH
• rostlinné proteiny MeSH

The preservation of enzymatic activity is a fundamental requirement for exploiting hybrid nano-bio-conjugates, and the control over protein-nanoparticle interactions, leading to stable and catalytically active hybrids, represents the key for designing new biosensing platforms. In this scenario, surface active maghemite nanoparticles (SAMNs) represent a new class of naked magnetic nanoparticles, displaying peculiar electrocatalytic features and the ability to selectively bind proteins. Recombinant aminoaldehyde dehydrogenase from tomato (SlAMADH1) was used as a model protein, and successfully immobilized by self-assembly on the surface of naked SAMNs, where its enzymatic activity resulted preserved for more than 6 months. The hybrid nanomaterial (SAMN@SlAMADH1) was characterized by UV-Vis spectroscopy, mass spectrometry, and TEM microscopy, and applied for the development of a biosensor for the determination of aminoaldehydes in alcoholic beverages. Measurements were carried out in a low volume electrochemical flow cell comprising a SAMN modified carbon paste electrode for the coulometric determination of the NADH produced during the enzymatic catalysis. The present findings, besides representing the first example of an electrochemical biosensor for aminoaldehydes in an alcoholic matrix, open the door to the use of immobilized enzymes on naked metal oxides nanomaterials for biosensing.

• Klíčová slova
• Aminoaldehyde biosensor, Aminoaldehyde dehydrogenase, Coulometric detection, Metal nanoparticles, NADH electro-oxidation, Nanomaterial electrocatalysis,
• MeSH
• aldehyddehydrogenasa metabolismus   MeSH
• aldehydy analýza   MeSH
• biosenzitivní techniky * MeSH
• elektrochemické techniky MeSH
• enzymy imobilizované metabolismus   MeSH
• kovové nanočástice chemie   MeSH
• propylaminy analýza   MeSH
• Solanum lycopersicum enzymologie   MeSH
• železité sloučeniny chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 3-aminopropionaldehyde MeSH Prohlížeč
• aldehyddehydrogenasa MeSH
• aldehydy MeSH
• enzymy imobilizované MeSH
• ferric oxide MeSH Prohlížeč
• propylaminy MeSH
• železité sloučeniny MeSH

• MeSH
• aldehydoxidoreduktasy metabolismus   MeSH
• aldehydy metabolismus   MeSH
• biogenní polyaminy metabolismus   MeSH
• buněčná stěna enzymologie   MeSH
• histocytochemie MeSH
• hrách setý metabolismus   MeSH
• indikátory a reagencie MeSH
• propylaminy metabolismus   MeSH
• semenáček MeSH
• tetrazoliová modř MeSH
• tkáňová distribuce MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• 3-aminopropionaldehyde MeSH Prohlížeč
• aldehydoxidoreduktasy MeSH
• aldehydy MeSH
• biogenní polyaminy MeSH
• gamma-guanidinobutyraldehyde dehydrogenase MeSH Prohlížeč
• indikátory a reagencie MeSH
• propylaminy MeSH
• tetrazoliová modř MeSH

Aldehyde dehydrogenases (ALDHs) constitute a superfamily of NAD(P)+-dependent enzymes, which detoxify aldehydes produced in various metabolic pathways to the corresponding carboxylic acids. Among the 19 human ALDHs, the cytosolic ALDH9A1 has so far never been fully enzymatically characterized and its structure is still unknown. Here, we report complete molecular and kinetic properties of human ALDH9A1 as well as three crystal forms at 2.3, 2.9, and 2.5 Å resolution. We show that ALDH9A1 exhibits wide substrate specificity to aminoaldehydes, aliphatic and aromatic aldehydes with a clear preference for γ-trimethylaminobutyraldehyde (TMABAL). The structure of ALDH9A1 reveals that the enzyme assembles as a tetramer. Each ALDH monomer displays a typical ALDHs fold composed of an oligomerization domain, a coenzyme domain, a catalytic domain, and an inter-domain linker highly conserved in amino-acid sequence and folding. Nonetheless, structural comparison reveals a position and a fold of the inter-domain linker of ALDH9A1 never observed in any other ALDH so far. This unique difference is not compatible with the presence of a bound substrate and a large conformational rearrangement of the linker up to 30 Å has to occur to allow the access of the substrate channel. Moreover, the αβE region consisting of an α-helix and a β-strand of the coenzyme domain at the dimer interface are disordered, likely due to the loss of interactions with the inter-domain linker, which leads to incomplete β-nicotinamide adenine dinucleotide (NAD+) binding pocket.

• Klíčová slova
• 3-aminopropionaldehyde, 4-trimethylaminobutyraldehyde, Homo sapiens, X‐ray crystallography, aldehyde dehydrogenase, structure-function,
• MeSH
• aldehyddehydrogenasa antagonisté a inhibitory   chemie   genetika   ultrastruktura   MeSH
• katalytická doména genetika   MeSH
• kinetika MeSH
• konformace proteinů * MeSH
• krystalografie rentgenová MeSH
• lidé MeSH
• NAD genetika   MeSH
• sekundární struktura proteinů MeSH
• sekvence aminokyselin genetika   MeSH
• substrátová specifita genetika   MeSH
• vazebná místa genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aldehyddehydrogenasa MeSH
• ALDH9A1 protein, human MeSH Prohlížeč
• NAD 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

Plant ALDH10 family members are aminoaldehyde dehydrogenases (AMADHs), which oxidize ω-aminoaldehydes to the corresponding acids. They have been linked to polyamine catabolism, osmoprotection, secondary metabolism (fragrance), and carnitine biosynthesis. Plants commonly contain two AMADH isoenzymes. We previously studied the substrate specificity of two AMADH isoforms from peas (PsAMADHs). Here, two isoenzymes from tomato (Solanum lycopersicum), SlAMADHs, and three AMADHs from maize (Zea mays), ZmAMADHs, were kinetically investigated to obtain further clues to the catalytic mechanism and the substrate specificity. We also solved the high resolution crystal structures of SlAMADH1 and ZmAMADH1a because these enzymes stand out from the others regarding their activity. From the structural and kinetic analysis, we can state that five residues at positions 163, 288, 289, 444, and 454 (PsAMADHs numbering) can, directly or not, significantly modulate AMADH substrate specificity. In the SlAMADH1 structure, a PEG aldehyde derived from the precipitant forms a thiohemiacetal intermediate, never observed so far. Its absence in the SlAMADH1-E260A structure suggests that Glu-260 can activate the catalytic cysteine as a nucleophile. We show that the five AMADHs studied here are capable of oxidizing 3-dimethylsulfoniopropionaldehyde to the cryo- and osmoprotectant 3-dimethylsulfoniopropionate. For the first time, we also show that 3-acetamidopropionaldehyde, the third aminoaldehyde besides 3-aminopropionaldehyde and 4-aminobutyraldehyde, is generally oxidized by AMADHs, meaning that these enzymes are unique in metabolizing and detoxifying aldehyde products of polyamine degradation to nontoxic amino acids. Finally, gene expression profiles in maize indicate that AMADHs might be important for controlling ω-aminoaldehyde levels during early stages of the seed development.

• MeSH
• aldehydoxidoreduktasy chemie   genetika   metabolismus   MeSH
• aldehydy chemie   MeSH
• chemické modely MeSH
• fylogeneze MeSH
• fyziologie rostlin MeSH
• kinetika MeSH
• krystalografie rentgenová metody   MeSH
• kukuřice setá enzymologie   MeSH
• mutageneze cílená MeSH
• NAD chemie   MeSH
• polyethylenglykoly chemie   MeSH
• regulace genové exprese enzymů * MeSH
• regulace genové exprese u rostlin * MeSH
• rostliny enzymologie   MeSH
• semena rostlinná metabolismus   MeSH
• Solanum lycopersicum enzymologie   MeSH
• substrátová specifita MeSH
• vazba proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aldehydoxidoreduktasy MeSH
• aldehydy MeSH
• NAD MeSH
• polyethylenglykoly MeSH

According to our knowledge, this is the first purification method developed, enabling isolation of a homogeneous aminoaldehyde dehydrogenase (AMADH) from etiolated pea seedlings. The procedure involved initial purification with precipitants followed by three low pressure chromatographic steps. Partially purified enzyme was further subjected to fast protein liquid chromatography on a Mono Q column and to affinity-interaction chromatography on 5'-AMP Sepharose. Purity of the final enzyme preparation was checked by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and chromatofocusing. Pea AMADH exists as a tetramer of 230 kDa in the native state, a molecular mass of one subunit was determined as 57 kDa. The enzyme was found to be an acidic protein with pI 5.4. AMADH showed a broad substrate specificity utilising various aminoaldehydes (C3-C6) as substrates. The best substrate of pea AMADH was 3-aminopropionaldehyde, the enzyme also efficiently oxidised 4-aminobutyraldehyde and omega-guanidinoanalogues of the aminoaldehydes. Pea AMADH was inhibited by SH reagents, several elementary aldehydes and metal-binding agents. Although AMADH did not oxidise betaine aldehyde at all, the N-terminal amino acid sequence of the enzyme shows a high degree of homology with those of plant betaine aldehyde dehydrogenases (BADHs) of spinach, sugar beet and amaranth. Several conserved amino acids were found in comparison with BADH from cod liver of known crystal structure.

• MeSH
• aldehydoxidoreduktasy antagonisté a inhibitory   chemie   izolace a purifikace   metabolismus   MeSH
• chromatografie kapalinová MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• hmotnostní spektrometrie MeSH
• hrách setý enzymologie   MeSH
• inhibitory enzymů farmakologie   MeSH
• koncentrace vodíkových iontů MeSH
• molekulární sekvence - údaje MeSH
• sekvence aminokyselin MeSH
• stabilita enzymů MeSH
• substrátová specifita MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aldehydoxidoreduktasy MeSH
• aminobutyraldehyde dehydrogenase MeSH Prohlížeč
• inhibitory enzymů MeSH