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PMID:12860978
| Citation |
Kupke, T, Hernández-Acosta, P and Culiáñez-Macià, FA (2003) 4'-phosphopantetheine and coenzyme A biosynthesis in plants. J. Biol. Chem. 278:38229-37 |
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| Abstract |
Coenzyme A is required for many synthetic and degradative reactions in intermediary metabolism and is the principal acyl carrier in prokaryotic and eukaryotic cells. Coenzyme A is synthesized in five steps from pantothenate, and recently the CoaA biosynthetic genes in bacteria and human have all been identified and characterized. Coenzyme A biosynthesis in plants is not fully understood, and to date only the AtHAL3a (AtCoaC) gene of Arabidopsis thaliana has been cloned and identified as 4'-phosphopantothenoylcysteine (PPC) decarboxylase (Kupke, T., Hernández-Acosta, P., Steinbacher, S., and Culiáñez-Macià, F. A. (2001) J. Biol. Chem. 276, 19190-19196). Here, we demonstrate the cloning of the four missing genes, purification of the enzymes, and identification of their functions. In contrast to bacterial PPC synthetases, the plant synthetase is not CTP-but ATP-dependent. The complete biosynthetic pathway from pantothenate to coenzyme A was reconstituted in vitro by adding the enzymes pantothenate kinase (AtCoaA), 4'-phosphopantothenoylcysteine synthetase (AtCoaB), 4'-phosphopantothenoylcysteine decarboxylase (AtCoaC), 4'-phosphopantetheine adenylyltransferase (AtCoaD), and dephospho-coenzyme A kinase (AtCoaE) to a mixture containing pantothenate, cysteine, ATP, dithiothreitol, and Mg2+. |
| Links |
PubMed Online version:10.1074/jbc.M306321200 |
| Keywords |
Adenosine Triphosphate/chemistry; Adenosine Triphosphate/metabolism; Amino Acid Sequence; Arabidopsis/chemistry; Arabidopsis/genetics; Biochemical Phenomena; Biochemistry; Cell Line; Cloning, Molecular; Coenzyme A/biosynthesis; Cysteine/chemistry; DNA, Complementary/metabolism; Electrophoresis, Polyacrylamide Gel; Escherichia coli/metabolism; Genome, Plant; Magnesium/chemistry; Models, Chemical; Molecular Sequence Data; Nucleotidyltransferases/chemistry; Nucleotidyltransferases/genetics; Nucleotidyltransferases/isolation & purification; Osmosis; Pantetheine/analogs & derivatives; Pantetheine/chemistry; Plants/metabolism; Recombinant Fusion Proteins/chemistry; Sequence Homology, Amino Acid; Time Factors |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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