Plant aminoaldehyde dehydrogenases oxidize a wide range of nitrogenous heterocyclic aldehydes
- 358 Downloads
The metabolic degradation of aldehydes is catalyzed by oxidoreductases from which aldehyde dehydrogenases (EC 1.2.1) comprise nonspecific or substrate-specific enzymes. The latter subset is represented, e.g., by NAD+-dependent aminoaldehyde dehydrogenases (AMADHs; EC 18.104.22.168) oxidizing a group of naturally occurring ω-aminoaldehydes including polyamine oxidation products. Recombinant isoenzymes from pea (PsAMADH1 and 2) and tomato (LeAMADH1 and 2) were subjected to kinetic measurements with synthetic aldehydes containing a nitrogenous heterocycle such as pyridinecarbaldehydes and their halogenated derivatives, (pyridinylmethylamino)-aldehydes, pyridinyl propanals and aldehydes derived from purine, 7-deazapurine and pyrimidine to characterize their substrate specificity and significance of the resulting data for in vivo reactions. The enzymatic production of the corresponding carboxylic acids was analyzed by liquid chromatography coupled to electrospray ionization mass spectrometry. Although the studied AMADHs are largely homologous and supposed to have a very similar active site architecture, significant differences were observed. LeAMADH1 displayed the broadest specificity oxidizing almost all compounds followed by PsAMADH2 and 1. In contrast, LeAMADH2 accepted only a few compounds as substrates. Pyridinyl propanals were converted by all isoenzymes, usually better than pyridinecarbaldehydes and aldehydes with fused rings. The K m values for the best substrates were in the range of 10−5–10−4 M. Nevertheless, the catalytic efficiency values (V max/K m) reached only a very small fraction of that with 3-aminopropanal (except for LeAMADH1 activity with two pyridine-derived compounds). Docking experiments using the crystal structure of PsAMADH2 were involved to discuss differences in results with position isomers or alkyl chain homologs.
KeywordsAldehyde Aminoaldehyde dehydrogenase 7-Deazapurine Pyridine Pyrimidine Purine
Betaine aldehyde dehydrogenase
Tomato (Lycopersicon esculentum) aminoaldehyde dehydrogenase
Liquid chromatography coupled to mass spectrometry
Pea (Pisum sativum) aminoaldehyde dehydrogenase
This work was supported by OP RD&I grant no. ED0007/01/01 (Centre of the Region Haná for Biotechnological and Agricultural Research) and grant no. MSM0021622413 from the Ministry of Education, Youth and Sports, Czech Republic, plus grant no. 522/08/0555 from the Czech Science Foundation. We would also like to thank Hana Moskalíková, a former student of the Department of Biochemistry, Faculty of Science, Palacký University, for her valuable contribution to initial experiments.
Conflict of interest
The authors declare no conflict of interest.
- Brocker C, Lassen N, Estey T, Pappa A, Cantore M, Orlova VV, Chavakis T, Kavanagh KL, Oppermann U, Vasiliou V (2010) Aldehyde dehydrogenase 7A1 (ALDH7A1) is a novel enzyme involved in cellular defense against hyperosmotic stress. J Biol Chem 285:18452–18463. doi: 10.1074/jbc.M109.077925 PubMedCrossRefGoogle Scholar
- Crews P, Rodríguez J, Jaspars M (2010) Organic structure analysis, 2nd edn. Oxford University Press, New York, p 273Google Scholar
- DeLano W (2002) The PyMOL molecular graphics system. DeLano Scientific, Palo Alto. http://www.pymol.org
- Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA, Cheeseman JR, Montgomery JA Jr, Vreven T, Kudin KN, Burant JC, Millam JM, Iyengar SS, Tomasi J, Barone V, Mennucci B, Cossi M, Scalmani G, Rega N, Petersson GA, Nakatsuji H, Hada M, Ehara M, Toyota K, Fukuda R, Hasegawa J, Ishida M, Nakajima T, Honda Y, Kitao O, Nakai H, Klene M, Li X, Knox JE, Hratchian HP, Cross JB, Bakken V, Adamo C, Jaramillo J, Gomperts R, Stratmann RE, Yazyev O, Austin AJ, Cammi R, Pomelli C, Ochterski JW, Ayala PY, Morokuma K, Voth GA, Salvador P, Dannenberg JJ, Zakrzewski VG, Dapprich S, Daniels AD, Strain MC, Farkas O, Malick DK, Rabuck AD, Raghavachari K, Foresman JB, Ortiz JV, Cui Q, Baboul AG, Clifford S, Cioslowski J, Stefanov BB, Liu G, Liashenko A, Piskorz P, Komaromi I, Martin RL, Fox DJ, Keith T, Al-Laham MA, Peng CY, Nanayakkara A, Challacombe M, Gill PMW, Johnson B, Chen W, Wong MW, Gonzalez C, Pople JA (2004) Gaussian 03, Revision C.02. Gaussian Inc., WallingfordGoogle Scholar
- Gruez A, Roig-Zamboni V, Grisel S, Salomoni A, Valencia C, Campanacci V, Tegoni M, Cambillau C (2004) Crystal structure and kinetics identify Escherichia coli YdcW gene product as a medium-chain aldehyde dehydrogenase. J Mol Biol 343:29–41. doi: 10.1016/j.jmb.2004.08.030 PubMedCrossRefGoogle Scholar
- Hill JP, Dickinson FM (1988) Pre-steady-state kinetics of aldehyde oxidation by pig liver cytosolic aldehyde dehydrogenase. Biochem Soc Trans 16:856–857Google Scholar
- Kopečný D, Tylichová M, Snégaroff J, Popelková H, Šebela M (2011) Carboxylate and aromatic active-site residues are determinants of high-affinity binding of ω-aminoaldehydes to plant aminoaldehyde dehydrogenases. FEBS J 278:3130–3139. doi: 10.1111/j.1742-4658.2011.08239.x PubMedCrossRefGoogle Scholar
- Morris GM, Goodsell DS, Halliday RS, Huey R, Hart WE, Belew RK, Olson AJ (1998) Automated docking using a Lamarckian genetic algorithm and an empirical binding free energy function. J Comput Chem 19:1639–1662. doi: 10.1002/(SICI)1096-987X(19981115)19:14<1639:AID-JCC10>3.0.CO;2-B CrossRefGoogle Scholar
- Pearlman DA, Case DA, Caldwell JW, Ross WS, Cheatham TE, Debolt S, Ferguson D, Seibel G, Kollman P (1995) Amber, a package of computer programs for applying molecular mechanics, normal mode analysis, molecular dynamics and free energy calculations to simulate the structural and energetic properties of molecules. Comput Phys Commun 91:1–41. doi: 10.1016/0010-4655(95)00041-D CrossRefGoogle Scholar
- Tylichová M, Briozzo P, Kopečný D, Ferrero J, Moréra S, Joly N, Snégaroff J, Šebela M (2008) Purification, crystallization and preliminary crystallographic study of a recombinant plant aminoaldehyde dehydrogenase from Pisum sativum. Acta Crystallogr Sect F Struct Biol Cryst Commun 64:88–90. doi: 10.1107/S1744309107068522 PubMedCrossRefGoogle Scholar
- Tylichová M, Kopečný D, Moréra S, Briozzo P, Lenobel R, Snégaroff J, Šebela M (2010) Structural and functional characterization of plant aminoaldehyde dehydrogenase from Pisum sativum with a broad specificity for natural and synthetic aminoaldehydes. J Mol Biol 396:870–882. doi: 10.1016/j.jmb.2009.12.015 PubMedCrossRefGoogle Scholar
- Warburg O, Christian W (1943) Isolierung und Kristallisation des Gärungsferments Zymohexase. Biochem Z 314:149–176 (in German)Google Scholar