Molecular cloning and functional identification of a plant ornithine decarboxylase cDNA
Molecular cloning and functional identification of a plant ornithine decarboxylase cDNA
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DOI：
10.1042/bj3140241
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发表时间：
1996-02-15
影响因子：
4.1
通讯作者：
Burtin, D
中科院分区：
文献类型：
--
作者：
Michael, AJ;Furze, JM;Burtin, D
从茄科植物曼陀罗（Datura stramonium）的根培养物中分离到一种植物鸟氨酸脱羧酶（ODC）的cDNA，该酶是腐胺和多胺生物合成的关键酶。逆转录-PCR采用简并寡核苷酸引物代表其他真核ODC的保守基序被用来分离的cDNA。最长的开放阅读框可能编码431个氨基酸的肽，并且表现出与其他真核ODC、原核和真核精氨酸脱羧酶（ADC）、原核内消旋二氨基庚二酸脱羧酶以及假单胞菌（Pseudomonasseringae cv.）烟粉虱参与小鼠ODC活性位点的残基在植物酶中是保守的。植物ODC不具有在哺乳动物酶中发现的C-末端延伸，涉及蛋白质的快速周转，这表明植物ODC可能具有较长的半衰期。在大肠杆菌中表达植物ODC并证明ODC活性证实cDNA编码活性ODC酶。这是第一次描述从存在腐胺替代ADC途径的生物体中分离的真核ODC的一级结构。
A cDNA for a plant ornithine decarboxylase (ODC), a key enzyme in putrescine and polyamine biosynthesis, has been isolated from root cultures of the solanaceous plant Datura stramonium. Reverse transcription-PCR employing degenerate oligonucleotide primers representing conserved motifs from other eukaryotic ODCs was used to isolate the cDNA. The longest open reading frame potentially encodes a peptide of 431 amino acids and exhibits similarity to other eukaryotic ODCs, prokaryotic and eukaryotic arginine decarboxylases (ADCs), prokaryotic meso-diaminopimelate decarboxylases and the product of the tabA gene of Pseudomonas syringae cv. tabaci. Residues involved at the active site of the mouse ODC are conserved in the plant enzyme. The plant ODC does not possess the C-terminal extension found in the mammalian enzyme, implicated in rapid turnover of the protein, suggesting that the plant ODC may have a longer half-life, Expression of the plant ODC in Escherichia coli and demonstration of ODC activity confirmed that the cDNA encodes an active ODC enzyme. This is the first description of the primary structure of a eukaryotic ODC isolated from an organism where the alternative ADC route to putrescine is present.