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

• Autorzy: Wisniewski P , Szklarczyk J. , Maciaga M. , Paszkowski A.
• Języki publikacji: EN

Abstrakty

EN

The occurrence of four L-alanine:2-oxoglutarate aminotransferase (AOAT) isoenzymes (AOAT-like proyeins): alanine aminotransferase 1 and 2 (AlaATl and AlaAT2, EC 2.6.1.2) and L-glutamate:glyoxylate aminotransferase 1 and 2 (GGAT1 and GGAT2, EC 2.6.1.4) 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.

Słowa kluczowe

EN

alanine aminotransferase; plant physiology; enzyme protein; Arabidopsis thaliana; isoenzyme; L-alanine:2-oxoglutarate aminotransferase; leaf

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2006
• Tom: 28
• Numer: 6
• Opis fizyczny: p.577-588,fig.,ref.

Twórcy

autor

Wisniewski P

• Warsaw Agricultural University, Nowoursynowska 159, 02-776 Warsaw, Poland

autor

Szklarczyk J.

autor

Maciaga M.

autor

Paszkowski A.

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