Pantothenoylcysteine‐4′‐phosphate Decarboxylase from Horse Liver

Scandurra, Roberto; Barboni, E.; Granata, Filippo; Pensa, Bernardo; Costa, Mara

doi:10.1111/j.1432-1033.1974.tb03805.x

Cited by 23 publications

(13 citation statements)

References 22 publications

(9 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The following qualitative experiments similar to those used by Hodgins and Abeles [5] to detect a pyruvoyl residue in the active site of an impure preparation of D-proline reductase demonstrate that pyruvate is present and functionally involved in horse liver phosphopantothenoylcysteine decarboxylase as suggested in [6,7].…”

Section: Introductionsupporting

confidence: 58%

“…Correspondence address: R. Scandurra, Dipartimento di Scienze Biochimiche, Universita 'La Sapienza', Ple Aldo Moro 5, 00185 Roma, Italy Horse liver phosphopantothenoylcysteine decarboxylase was prepared as described [6,7]. The enzyme had a specific activity of about 80 nmol CO2 .…”

Section: Methodsmentioning

confidence: 99%

“…The samples were then diluted with water and exhaustively dialyzed against 0.05 M phosphate buffer (pH 6.5): proteins, radioactivity and enzyme activity were then measured. A parallel experiment in the presence of 15 mM cold substrate (lo-times its K,,, value [6]) was conducted (0).…”

Section: Tritium Incorporation Into the Proteinmentioning

confidence: 99%

See 2 more Smart Citations

Covalently bound pyruvate in phosphopantothenoylcysteine decarboxylase from horse liver

Scandurra¹,

Politi²,

Santoro³

et al. 1987

FEBS Letters

View full text Add to dashboard Cite

Horse liver phosphopantothenoylcysteine decarboxylase (EC 4.1.1.36) incorporates nonexchangeable tritium from borotritide with a decrease of the activity. Substrate prevents both tritium incorporation and the decrease in activity. Acid and base hydrolysis of the tritiated protein releases labeled lactate identified by highvoltage paper electrophoresis, paper chromatography and silicic acid chromatography. These results indicate the presence of pyruvate covalently bound through an ester linkage to phosphopantothenoylcysteine decarboxylase which is then another example of a mammalian enzyme in which pyruvate is involved in a catalytic activity.

show abstract

Section: Introductionsupporting

confidence: 58%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Covalently bound pyruvate in phosphopantothenoylcysteine decarboxylase from horse liver

Scandurra¹,

Politi²,

Santoro³

et al. 1987

FEBS Letters

View full text Add to dashboard Cite

show abstract

“…Conclusion-The Dfp protein from E. coli bears little resemblance to PPC-S and PPC-DC from mammalian sources: PPC-DC has been partially purified from both horse (17) and rat liver (18), and in the case of the former, it was shown that the enzyme is pyruvoyl-dependent (19). PPC-S from rat liver uses ATP as the activating nucleotide, instead of CTP, and forms an acyl-phosphate intermediate as suggested by the formation of ADP and phosphate (16).…”

Section: Purification Of the Native Protein And Identification Of Thementioning

confidence: 99%

Phosphopantothenoylcysteine Synthetase from Escherichia coli

Strauss

Kinsland

et al. 2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

Phosphopantothenoylcysteine synthase catalyzes the formation of (R)-4-phospho-N-pantothenoylcysteine from 4-phosphopantothenate and L-cysteine: this enzyme, involved in the biosynthesis of coenzyme A (CoA), has not previously been identified. Recently it was shown that the NH 2 -terminal domain of the Dfp protein from bacteria catalyzes the next step in CoA biosynthesis, the decarboxylation of (R)-4-phospho-N-pantothenoylcysteine to form 4-phosphopantetheine (Kupke, T., Uebele, M., Schmid, D., Jung, G., Blaesse, M., and Steinbacher, S. (2000) J. Biol. Chem. 275, 31838 -31846). We have partially purified phosphopantothenoylcysteine decarboxylase from Escherichia coli and demonstrated that the protein encoded by the dfp gene, here renamed coaBC, also has phosphopantothenoylcysteine synthetase activity, using CTP rather than ATP as the activating nucleoside 5-triphosphate. This discovery completes the identification of all the enzymes involved in the biosynthesis of coenzyme A in bacteria.Coenzyme A (CoA) 1 fulfils a vital role in the normal metabolism of living organisms: it has been reported that ϳ4% of all enzymes utilize either CoA, its thioesters or 4Ј-phosphopantetheine (1). As an essential cofactor of fatty acid synthases, polyketide synthases, and nonribosomal peptide synthases, 4Ј-phosphopantetheine functions as an acyl group carrier, activating carboxylic acids for biological Claisen reactions and the formation of peptides and esters. Acetyl-CoA and succinyl-CoA are key intermediates in energy metabolism, taking part in the tricarboxylic acid cycle. It is therefore surprising that three of the CoA biosynthetic enzymes have only recently been identified in E. coli (2,3).Phosphopantothenoylcysteine decarboxylase (PPC-DC) catalyzes the decarboxylation of 4Ј-phosphopantothenoylcysteine (PPanCys) to form 4Ј-phosphopantetheine (Fig. 1). From a mechanistic perspective this reaction proves to be particularly interesting as it is unclear how the stabilization of the carbanion intermediate is achieved. To study this enzyme we have repeated, with major modifications, the published purification of PPC-DC from wild-type E. coli (4). Amino-terminal sequencing identified the dfp gene as coding for the protein having this activity. The flavin-containing Dfp protein was first identified from a temperature-sensitive mutant (dfp-707), believed to be involved in DNA and pantothenate metabolism (5, 6). However, the conditional lethality of the mutant was reported not to be a direct result of pantothenate auxotrophy, and its complete characterization was not achieved. While our work was in progress, Kupke et al. (7) also identified Dfp as having PPC-DC activity by sequence comparison with EpiD, a lantibiotic-synthesizing protein catalyzing the decarboxylation of peptidylcysteine intermediates.We now report that the protein encoded by dfp also catalyzes the formation of PPanCys from 4Ј-phosphopantothenate and L-cysteine using cytidine 5Ј-triphosphate (CTP) as the activating nucleoside 5Ј-triphosphate. The protein releases CM...

show abstract

“…However, such a complex has not been observed in other organisms. In mammalian cells, the first three enzymes in the pathway of CoA biosynthesis are cytosolic proteins (17,22,23). The data on the localization of CoA synthase are somewhat controversial.…”

mentioning

confidence: 99%

Subcellular Localization and Regulation of Coenzyme A Synthase

Zhyvoloup

Nemazanyy

Panasyuk

et al. 2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

CoA synthase mediates the last two steps in the sequence of enzymatic reactions, leading to CoA biosynthesis. We have recently identified cDNA for CoA synthase and demonstrated that it encodes a bifunctional enzyme possessing 4-phosphopantetheine adenylyltransferase and dephospho-CoA kinase activities. Molecular cloning of CoA synthase provided us with necessary tools to study subcellular localization and the regulation of this bifunctional enzyme. Transient expression studies and confocal microscopy allowed us to demonstrate that full-length CoA synthase is associated with the mitochondria, whereas the removal of the Nterminal region relocates the enzyme to the cytosol. In addition, we showed that the N-terminal sequence of CoA synthase (amino acids 1-29) exhibits a hydrophobic profile and targets green fluorescent protein exclusively to mitochondria. Further analysis, involving subcellular fractionation and limited proteolysis, indicated that CoA synthase is localized on the mitochondrial outer membrane. Moreover, we demonstrate for the first time that phosphatidylcholine and phosphatidylethanolamine, which are the main components of the mitochondrial outer membrane, are potent activators of both enzymatic activities of CoA synthase in vitro. Taken together, these data provide the evidence that the final stages of CoA biosynthesis take place on mitochondria and the activity of CoA synthase is regulated by phospholipids.

show abstract

Pantothenoylcysteine‐4′‐phosphate Decarboxylase from Horse Liver

Cited by 23 publications

References 22 publications

Covalently bound pyruvate in phosphopantothenoylcysteine decarboxylase from horse liver

Covalently bound pyruvate in phosphopantothenoylcysteine decarboxylase from horse liver

Phosphopantothenoylcysteine Synthetase from Escherichia coli

Subcellular Localization and Regulation of Coenzyme A Synthase

Contact Info

Product

Resources

About