Identification of a second pyridoxine (pyridoxamine) 5′-phosphate oxidase in Arabidopsis thaliana

Autor: Narendra Singh, Leslie R. Goertzen, Ywh-Min Tzou, Robert D. Locy, Yuying Sang
Rok vydání: 2010
Předmět:
Zdroj: Acta Physiologiae Plantarum. 33:559-566
ISSN: 1861-1664
0137-5881
DOI: 10.1007/s11738-010-0579-6
Popis: Pyridoxine (pyridoxamine) 5′-phosphate oxidase (PPOX) is involved in the biosynthetic pathway of vitamin B6, converting pyridoxine 5′-phosphate (PNP) or pyridoxamine 5′-phosphate (PMP) into pyridoxal 5′-phosphate (PLP). PLP is a well-known cofactor of numerous enzymes including transamination and decarboxylation reactions. We have previously identified a PPOX (AtPPOX-1) protein encoded by At5g49970 in Arabidopsis thaliana. Here, we report a second PPOX in Arabidopsis, which was named as AtPPOX-2 encoded by At2g46580. The RT-PCR amplified cDNA of AtPPOX-2 was cloned into an Escherichia coli expression vector and a yeast shuttle vector. Both PPOX enzyme assay and complementation of the oxidative stress sensitivity phenotype of a yeast PDX3 deletion mutant demonstrated that At2g46580 encodes a PPOX protein (AtPPOX-2). The catalytic efficiency of AtPPOX-1 is approximately 300-fold higher than that of AtPPOX-2 for PNP. Based on bioinformatic analysis, AtPPOX-2 has a putative mitochondrial transit peptide at the N-terminus. The truncated AtPPOX-2 without 18 amino acids at the N-terminal end lost PPOX activity, suggesting that the N-terminal 18 amino acids are necessary for the enzyme activity of AtPPOX-2. Phylogenetic analysis of AtPPOX-2 homologs from all domains of life suggests that AtPPOX-2 homologs in plants are the product of lateral gene transfer from the cyanobacterial endosymbionts from which plastids are derived.
Databáze: OpenAIRE
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