Cofactor Biosynthesis: A Mechanistic Perspective

Begley, Tadhg P.; Kinsland, Cynthia; Taylor, Sean V.; Tandon, Manish; Nicewonger, Robb; Wu, Min; Chiu, Hsiu‐Ju; Kelleher, Neil L.; Campobasso, Nino; Zhang, Yi

doi:10.1007/3-540-69542-7_3

Cited by 28 publications

(28 citation statements)

References 182 publications

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“…Identification of the dephospho-CoA kinase gene provides a major missing piece of information in the genetics of CoA biosynthesis. Of the nine enzymes required for CoA biosynthesis, only the gene for phosphopantothenoyl cysteine synthetase has not been identified yet (3). Identification of homologues of coaE should facilitate further studies of CoA biosynthesis in other organisms.…”

Section: Discussionmentioning

confidence: 99%

“…The final two steps in CoA biosynthesis are coupling of phosphopantetheine with ATP to form dephosphocoenzyme A (dephospho-CoA) and the subsequent phosphorylation of the 3Ј-hydroxyl group to form CoA ( Fig. 1) (3). Genes coding for six of the seven enzymes involved in the biosynthesis of phosphopantetheine have been identified (10,14,20,26).…”

mentioning

confidence: 99%

“…Enzyme assays and product characterization were used to show that the resulting recombinant protein is indeed dephosphoCoA kinase. The dephospho-CoA kinase gene is now designated coaE, based on the previous naming of the pantothenate kinase gene as coaA and of the phosphopantetheine adenylyltransferase gene as coaD (3,7).…”

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confidence: 99%

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Identification of yacE ( coaE ) as the Structural Gene for Dephosphocoenzyme A Kinase in Escherichia coli K-12

2001

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Dephosphocoenzyme A (dephospho-CoA) kinase catalyzes the final step in coenzyme A biosynthesis, the phosphorylation of the 3-hydroxy group of the ribose sugar moiety. Wild-type dephospho-CoA kinase from Corynebacterium ammoniagenes was purified to homogeneity and subjected to N-terminal sequence analysis. A BLAST search identified a gene from Escherichia coli previously designated yacE encoding a highly homologous protein. Amplification of the gene and overexpression yielded recombinant dephospho-CoA kinase as a 22.6-kDa monomer. Enzyme assay and nuclear magnetic resonance analyses of the product demonstrated that the recombinant enzyme is indeed dephospho-CoA kinase. The activities with adenosine, AMP, and adenosine phosphosulfate were 4 to 8% of the activity with dephospho-CoA. Homologues of the E. coli dephospho-CoA kinase were identified in a diverse range of organisms.Coenzyme A (CoA) is an essential cofactor in a wide variety of biochemical pathways (1). It is estimated that about 4% of all enzymes use CoA or a thioester of CoA as a substrate (24). The final two steps in CoA biosynthesis are coupling of phosphopantetheine with ATP to form dephosphocoenzyme A (dephospho-CoA) and the subsequent phosphorylation of the 3Ј-hydroxyl group to form CoA ( Fig. 1) (3). Genes coding for six of the seven enzymes involved in the biosynthesis of phosphopantetheine have been identified (10,14,20,26). The final two reactions of CoA biosynthesis in mammalian cells were first attributed to a single bifunctional enzyme by Hoagland and Novelli (8). This was verified by Worrall and Tubbs, who purified a protein from pork liver that possessed both the phosphopantetheine adenylyltransferase and dephospho-CoA kinase activities (25). Subsequently, these two activities were shown to be part of a multifunctional enzyme complex in baker's yeast (5, 6). We first showed that the final two steps in CoA biosynthesis in Corynebacterium ammoniagenes (formerly Brevibacterium ammoniagenes) are catalyzed by distinct proteins that were readily separated by ion-exchange chromatography (12). The gene from Escherichia coli coding for phosphopantetheine adenylyltransferase was recently cloned, and the crystal structure of the enzyme was determined (7, 9). We report here purification of dephospho-CoA kinase from wildtype C. ammoniagenes and identification of the gene coding for a homologous protein in E. coli. This gene was cloned using PCR and overexpressed, and the resulting protein was purified. Enzyme assays and product characterization were used to show that the resulting recombinant protein is indeed dephosphoCoA kinase. The dephospho-CoA kinase gene is now designated coaE, based on the previous naming of the pantothenate kinase gene as coaA and of the phosphopantetheine adenylyltransferase gene as coaD (3, 7). MATERIALS AND METHODSMaterials. Dephospho-CoA, dilithium CoA, NADH, pyruvate kinase, lactic dehydrogenase, phosphoenolpyruvate, ATP, AMP, adenosine phosphosulfate (APS), DEAE Sepharose, and Q Sepharose were purchased from Sigma. A Sep...

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Section: Discussionmentioning

confidence: 99%

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Identification of yacE ( coaE ) as the Structural Gene for Dephosphocoenzyme A Kinase in Escherichia coli K-12

2001

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“…The biosynthesis of the thiazole moiety (2) of thiamin involves the oxidative condensation of deoxy-D-xylulose-5-phosphate (1), tyrosine, and cysteine (Scheme 1) (1)(2)(3)(4).…”

Section: T Hiamin Pyrophosphate (3) Is An Essential Cofactor In All Lmentioning

confidence: 99%

Biosynthesis of the thiazole moiety of thiamin in Escherichia coli : Identification of an acyldisulfide-linked protein–protein conjugate that is functionally analogous to the ubiquitin/E1 complex

Kinsland

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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109

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A covalently linked protein-protein conjugate between ThiF and ThiS thiocarboxylate was found in a partially purified coexpressed ThiF͞ThiS protein mixture by using Fourier transform mass spectrometry. The Cys-184 of ThiF and the C terminus of ThiS thiocarboxylate were identified to be involved in the formation of this complex by using both mutagenesis and chemical modification methods. A complementation study of Escherichia coli thiF ؊ using thiF(C184S) suggests that this conjugate is an essential intermediate involved in the biosynthesis of the thiazole moiety of thiamin. This ThiF͞ThiS conjugate is the first characterized example of a unique acyldisulfide intermediate in a biosynthetic system. This protein conjugate is also an example of an ubiquitin-E1 like protein-protein conjugate in prokaryotes and supports a strong evolutionary link between thiamin biosynthesis and the ubiquitin conjugating system.

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“…In nature the K vitamins are synthesised via the shikimic acid pathway, which is also utilised in the production of the aromatic amino acids [7]. The various prenyl side chains are synthesised via the terpene biosynthetic pathway before incorporation into the final compound [8]. Vitamin K 1 is synthesised by plants and is found at significant levels in green leafy vegetables, certain legumes and some vegetable oils, such as rapeseed and soyabean oil [9].…”

Section: Synthesis Of Naturally Occurring K Vitaminsmentioning

confidence: 99%