Preferencje help
Widoczny [Schowaj] Abstrakt
Liczba wyników
2006 | 28 | 6 |
Tytuł artykułu

L-alanine:2-oxoglutarate aminotransferase isoenzymes from Arabidopsis thaliana leaves

Warianty tytułu
Języki publikacji
The occurrence of four L-alanine:2-oxoglutarate aminotransferase (AOAT) isoenzymes (AOAT-like proyeins): alanine aminotransferase 1 and 2 (AlaATl and AlaAT2, EC and L-glutamate:glyoxylate aminotransferase 1 and 2 (GGAT1 and GGAT2, EC was demonstrated in Arabidopsis thaliana leaves. These enzymes differed in their substrate specificity, susceptibility to pyridoxal phosphate inhibitors and behaviour during molecular sieving on Zorbax SE-250 column. A difference was observed in the electrostatic charge values at pH 9.1 between GGAT1 and GGAT2 as well as between AlaAT1 and AlaAT2, despite high levels of amino acid sequence identity (93 % and 85 %, respectively). The unprecedented evidence for the monomeric structure of both AlaAT1 and AlaAT2 is presented. The molecular mass of each enzyme estimated by molecular sieving on Sephadex G-150 and Zorbax SE-250 columns and SDS/PAGE was approx-mately 60 kDa. The kinetic parameters: Km (Ala) = 1.53 mM, Km (2-oxoglutarate) = 0.18 mM, kcat = 124.6 s⁻¹, kcat/Km = 8.1 x 10⁴ M⁻¹-s⁻¹ of AlaAT1 were comparable to those determined for other AlaATs iso-ated from different sources. The two studied GGATs also consisted of a single subunit with molecular mass of 47.3-70 kDa. The estimated Km values for L-glutamate (1.2 mM) and glyoxylate (0.42 mM) in the transamination catalyzed by putative GGAT1 contributed to indentification of the enzyme. Based on these results we concluded that each of four AOAT genes in Arabidopsis thaliana leaves expresses different AOAT isoenzyme, functioning in a native state as a monomer.
Opis fizyczny
  • Warsaw Agricultural University, Nowoursynowska 159, 02-776 Warsaw, Poland
  • Andrews P. 1965. The gel-filtration behaviour of proteins related to their molecular weights over a wide range. Biochem. J., 96: 595-605.
  • Biekmann S., Feierabend J. 1982. Subcellular distribution, multiple forms and development of glutamate-pyruvate (glyoxylate) aminotransferase in plant tissues. Biochim. Biophys. Acta, 721: 268-279.
  • Blum H., Bayer H., Gross H.J. 1987. Improved silver staimng of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8: 93-99.
  • Bradford M.M. 1976. A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72: 248-254.
  • Chapman K.S.R., Hatch M.D. 1981. Aspartate decarboxylation in bundle sheath cells of Zea mays and its possible contribution to C4 photosynthesis. Aust. J. Plant. Physiol., 8: 237-248.
  • Givan C.V. 1980. Aminotransferases in higher plants. In: Biochemtstry of plants, ed. by Stumpf P.K., Conn E.E., Academic Press: 329-357.
  • Good A.G., Muench D.G. 1992. Purification and characterization of an anaerobically induced alanine aminotransferase from barley roots. Plant Physiol., 99: 1520-1525.
  • Grishin N.V., Phillips M.A., Golds mith E.J. 1995. Modelmg of spatial structiire of eukaryotic ornithine decarboxylases. Protein Science, 4: 1291-1304.
  • Hath M.D., Mau S.L. 1972. Activity, location and role of aspartate aminotransferase isoenzymes in leaves with C4 pathway photosynthesis. Arch. Biochem. Biophys., 156: 195-206.
  • Horder M., Rej R. 1983 . Alanine aminotransferase. In: Methods of enzymatic analysis, ed. by H.U. Bergmeyer, Verlag Chemie: 444-456.
  • Igarashi D., Miwa T., Seki M., Kobayashi M., Kato T., Tabata S., Shinozaki K., Ohsumi C. 2003. Identification of photorespiratory glutamate:glyoxylate aminotransferase (GGAT) gene in Arabidopsis. The Plant Journal, 33: 975-987.
  • Kikuchi H., Hirose S., Toki S., Kazuhito A., Takaiwa F.1999. Molecular characterization of a gene for alanine aminotransferase from rice (Oryza sativa). Plant Mol. Biol., 39: 149-159.
  • Koshiba T., Mito N., Miyakado M. 1993. L- and D-Tryptophan aminotransferases from maize coleoptiles. J. Plant Res., 106: 25-29.
  • Laemmli U.K. 1970. Cleavage of structiiral p^rtems during the assembly of the head of bacteriophage T4. Nature, 227: 680-685.
  • Lain-Guelbenzu B., Cardenas J., Munoz-Blanco J. 1991. Purification and properties of L-alanine aminotransferase from Chlamydomonas reinhardtii. Eur. J. Biochem., 202: 881-887.
  • Lee I.S., Muragaki Y., Ideguchi T., Hase T., Tsuji M., Ooshima A., Okuno E., Kido R. 1995. Molecular cloning and sequencing of cDNA enrodiig alanine-glyroxylate aminotransferase 2 from rat kidney. J. Biochem., 117: 856-862.
  • Leegood R.C., Lea P.J., Adcook M.D., Hausler R. 1995. The regulation and control of photorespiration. J. Exp. Bot., 46: 1397-1411.
  • Liepman A.H., Olsen L.J. 2001. Peroxisomal alanine: glyoxylate aminotransferase (AGT1) is a photorespiratory enryme with multiple substrates in Arabidopsis thaliana. The Plant Journal, 25: 487-498.
  • Liepman A.H., Olsen L.J. 2003. Alanine aminotransferase homologs catalyze the glutamate:glyoxylate aminotransferase reaction in peroxisomes of Arabidopsis. Plant Physiol., 131: 215-227.
  • Liepman A.H., Olsen L.J. 2004. Genomic analysis of aminotransferases in Arabidopsis thaliana. Critical Reviews in Plant Sciences, 23: 73-89.
  • Morino Y., Tanase S. 1985. Quasisubstrates and irreversible inhibitors of aspartate aminotransferase. In: Transaminases, ed. by Christen P., Metzler D.E., John Wiley and sons: 251-265.
  • Muench D.G., Christopher M.E., Good A.G. 1998. Cloning and expression of a hypoxic and nitrogen inducible maize alanine aminotransferase gene. Physiol Plant, 103: 503-512.
  • Murashige T., Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plantarum, 15: 473-497.
  • Noguchi T., Hayashi S. 1981. Plant leaf alanine: 2-oxoglutarate aminotransferase. Biochem. J., 195: 235-239.
  • Ohnishi J., Yamazaki M., Kanai R. 1983. Differentiation of photorespiratory activity between mesophyl and bundlle sheath cells of C4 plants II. Peroxisomes of Panicum miliaceum L. Plant Cell Physiol., 26: 797-803.
  • Otter T., Penther J.M., Mohr H. 1992. Control of the appearance of alanine aminotransferase in the Scots pine (Pinus sylvestris L.) seedling. Planta, 188: 376-383.
  • Orzechowski S., Socha-Hanc J., Paszkowski A. 1999a. Alanine aminotransferase and glycine amino a transferase from maize (Zea mays L.) leaves. Acta Biochim. Polon., 46: 447-457.
  • Orzechowski S., Socha-Hanc J., Paszkowski A. 1999b. Purification and properties of alanine aminotransferase from maize (Zea mays L.) leaves. Acta Physiol. Plant., 21: 323-330.
  • Orzechowski S., Socha-Hanc J., Paszkowski A. 1999c. Subcellular dist ribut ion of alanine aminotransferase from maize (Zea mays L.) leaves. Acta Physiol. Plant., 21: 331-334.
  • Paszkowski A., Niedzielska A. 1989. Glutamate:- glyoxylate aminotransferase from the seedtings of rye (Secale cereale L.). Acta Biochim. Polon., 36: 17-29.
  • Penther J.M. 1991. Analysis of alanine and aspartate aminotransferase isoforms in mustard (Sinapis alba L.) cotyledons. J. Chromatogr., 587: 101-108.
  • Price N.C., Stevens L. 1999. Fundamentals of enzymology. Oxtord Univeraity Press, Oxtord, New York: 118-153.
  • Rech J., Crouzet J. 1974. Partial purification and initial studi es of the tomato L-alanine-2-oxoglutarate aminotransferase. Biochim. Biophys. Acta, 350: 392-399.
  • Rowsell E.W., Carnie J.A., Snell K., Tatak B. 1972. Assays for glyoxylate aminotransferase activities. Int. J. Biochem., 3: 247-257.
  • Schneider G., Kack H., Lindqvist Y. 2000. The manifold of vitamin B6 dependent enzymes. Structure, 8: 1-6.
  • Sechley K.A., Yamaya T., Oaks A. 1992. Compart- mentation of nitrogen assimilation in higher plants. Int. Rev. Cyt., 136: 85-163.
  • Sheehan D., FitzGerald R. 1996. Ion exchange chromatography. In: Protein purification protocols, ed. by Doonan S., Humana Press: 145-150.
  • Simpson R.M., Nonhebel H.M., Christie D.L. 1997. Partial purification and characterization of an aromatic amino acid aminotransferase from mung bean (Vigna radiata L. Wilczek). Planta, 201: 71-77.
  • Son D., Jo J., Sugiyama T. 1991. Purification and characterization of alanine aminotransferase from Panicum miliaceum leaves. Archiv. Biochem. Biophys., 289: 262-266.
  • Son D., Sugiyama T. 1992. Molecular cloning of an alanine aminotransferase from NAD-malic enzyme type C4 plant Panicum miliaceum. Plant. Mol. Biol., 20: 705-713.
  • Taler D., Galperin M., Benjamin I., Cohen Y., Kenigsbuch D. 2004. Plant eR genes that enc ode photorespiratory enzymes confer resistance against disease. Plant Cell, 16: 172-184.
  • Truszkiewicz W., Paszkowski A. 2004. Serine:glyoxylate aminotransferases from maize and wheat leaves: purification and properties. Photosynthesis Res., 82: 35-47.
  • Ward D.E., Kengen S.W.M., van der Oost J., de Vos W.M. 2000. Purification and characterization of the alanine aminotransferase from hyperthermophilic archaeon Pyrococcusfuriosus and its role in alanine production. J. Bacteriol., 182: 2559-2566.
  • Wint er H., Robi ns on D.G., Heldt H.W. 1994. Subcellular volumes and metabolite concentrations in spinach leaves. Planta, 193: 530-535.
Typ dokumentu
Identyfikator YADDA
JavaScript jest wyłączony w Twojej przeglądarce internetowej. Włącz go, a następnie odśwież stronę, aby móc w pełni z niej korzystać.