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Journal of Bacteriology, May 2001, p. 2774-2778, Vol. 183, No. 9
Department of Chemistry, State University at
Stony Brook, Stony Brook, New York
Received 29 November 2000/Accepted 6 February 2001
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 3'-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 wild-type
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 dephospho-CoA 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).
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.9.2774-2778.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Identification of yacE (coaE)
as the Structural Gene for Dephosphocoenzyme A Kinase in
Escherichia coli K-12
ABSTRACT
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
INTRODUCTION
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
View larger version (23K):
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FIG. 1.
Final two steps of CoA biosynthesis (coaD,
phosphopantetheine adenylyltransferase; coaE, dephospho-CoA kinase).
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MATERIALS AND METHODS |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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Materials. 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 Sephacryl S-200 gel filtration column was purchased from Pharmacia. E. coli W3110 and C. ammoniagenes ATCC 6871 were obtained from the American Type Culture Collection. PCR primers were purchased from Integrated DNA Technologies, and the pEt 28b(+) vector was purchased from Novagen.
Enzyme and protein assays.
All enzyme assays were carried
out at 25°C, and changes in absorbance at 340 nm were monitored. One
unit of activity corresponded to the formation of 1 µmol of
product/min using a molar absorbtivity for NADH of 6,220 M
1 cm1. Dephospho-CoA kinase activity was
measured as described previously (12). Each 1-ml assay
mixture contained KCl (20 mM), MgCl2 (10 mM), ATP (10 mM),
NADH (0.3 mM), phosphoenolpyruvate (0.4 mM), pyruvate kinase (10 U),
and lactic dehydrogenase (4 U) in 50 mM Tris buffer (pH 8.5). After
addition of the dephospho-CoA kinase sample, the reaction was initiated
by adding dephospho-CoA (0.4 mM). Adenosine, APS, or AMP (0.4 mM) was
substituted for dephospho-CoA in assays performed with these alternate
substrates. To assay for CoA formation, each 1-ml assay mixture
contained MgCl2 (10 mM), ATP (10 mM), acetyl phosphate (0.4 mM), phosphate acetyltransferase (2 U), and a dephospho-CoA kinase
sample (0.02 U). The reaction was initiated by adding dephospho-CoA
(0.4 mM), and thioester formation was observed at 240 nm
(
240 = 5.4 × 103 M1
cm1). Concentrations of dephospho-CoA solutions were
measured by end point assays by using dephospho-CoA kinase. Kinetic
parameters were determined by fitting the data to the equation for a
sequential reaction mechanism by using Origin 50. For proof of product
structure, a solution containing dephospho-CoA (2.2 mg, 3.0 µmol),
ATP (0.1 µmol), MgCl2 (1.0 µmol), KCl (1.0 µmol), and
phosphoenolpyruvate (3.5 µmol) in 5 ml of H2O was
adjusted to pH 8.0, pyruvate kinase (5 U) and recombinant dephospho-CoA
kinase (2 U) were added, and the solution was left at room temperature
for 30 min. The CoA produced was purified by high-performance liquid
chromatography on a preparative C18 column (21.4 mm by 25 cm; 5 min with 5% methanol in 10 mM phosphate buffer [pH 4.5],
followed by a linear gradient to 40% methanol over 30 min) and had an
elution time of 20 min. The product was lyophilized and redissolved in
D2O for nuclear magnetic resonance (NMR) analysis. Protein
determinations were carried out by the method of Bradford
(4), using bovine serum albumin as the standard. Sodium
dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was
carried out with 12% polyacrylamide gels by using Tris-glycine buffer
(11).
Purification of wild-type dephospho-CoA kinase. C. ammoniagenes was grown at 26°C to an optical density at 650 nm of 1.7 in 2 liters of liquid medium containing (per liter) 10 g of glucose, 15 g of peptone, 3 g of dipotassium hydrogen phosphate, 2 of g of sodium chloride, 1 g of yeast extract, and 0.1 g of anhydrous magnesium sulfate. All subsequent steps were performed at 4°C. Cells were harvested by centrifugation, and the wet cell paste (16.9 g) was suspended in Tris buffer (50 mM Tris [pH 8.0], 5 mM dithiothreitol [DTT], 0.2 mM phenylmethylsulfonyl fluoride) to give a total volume of 60 ml. Cells were disrupted with a French press, and cellular debris was removed by centrifugation (10,000 × g, 20 min).
The supernatant (about 50 ml) was loaded onto a DEAE Sepharose column (2.5 by 10 cm) which had been preequilibrated with DEAE buffer (25 mM Tris [pH 8.0], 5 mM DTT). The column was washed with DEAE buffer containing NaCl (0.2 M). The column was eluted with a 300-ml linear NaCl gradient (0.2 to 0.5 M NaCl) in DEAE buffer. Dephospho-CoA kinase activity eluted at 0.37 M NaCl. The active fractions were pooled (11 ml), and MgCl2 was added to a final concentration of 0.01 M. The resulting combined fraction was then applied to a Red A agarose column (1.5 by 4.5 cm) which had been equilibrated with Red A agarose buffer (0.02 M Tris [pH 8.0], 0.01 M MgCl2, 2 mM DTT). The enzyme was allowed to bind to the Red A agarose for 35 min, and the column was washed with Red A agarose buffer (20 ml). The column was washed with the same buffer (15 ml) containing ATP (2 mM), and then the column was allowed to stand for 1 h. Dephospho-CoA kinase was eluted from the column with Red A agarose buffer (15 ml) containing ATP (2 mM). The active fractions (6 ml) were combined and applied to a Q Sepharose column (1.5 by 2.5 cm) equilibrated with Q buffer (50 mM Tris [pH 7.5], 2 mM DTT). The column was washed with Q buffer (10 ml) and eluted with a gradient of the same buffer (100 ml) containing KCl (0 to 0.5 M). Dephospho-CoA kinase activity eluted at 0.32 M KCl. The active fractions were pooled and concentrated to 2 ml. A portion of the resulting sample was submitted for N-terminal sequence analysis.Cloning and expression of the coaE gene. The coaE gene was amplified by using E. coli W3110 genomic DNA as the template in a PCR. The forward primer included an Ala codon (GCC) inserted after the start codon to form an NcoI restriction site: GGA ACC ATG GCC AGG TAT ATA GTT GCC TTA. The reverse primer included an HindIII restriction site after the stop codon: GCG CAA GCT TAC GGT TTT TCC TGT GAG ACA. The amplification cycle was as follows: denaturation at 94°C for 45 s, annealing at 58°C for 45 s, and extension at 72°C for 2 min. Amplification was performed for 32 cycles and was followed by a final extension step at 72°C for 10 min. The resulting PCR product was subcloned into the NcoI-HindIII site of the pEt28b(+) expression plasmid. The resulting plasmid, designated pEt/coaE, was sequenced by using the T7 promoter primer. E. coli BL21(DE3) was transformed with pEt28b/coaE to obtain an overexpression strain. All steps were carried out by using standard molecular biology procedures (16).
Purification of recombinant dephospho-CoA kinase.
E.
coli BL21(DE3)(pEt28b/coaE) was grown at 37°C in
Luria-Bertani medium (500 ml) containing kanamycin (50 µg/ml).
Isopropyl-
-D-thiogalactopyranoside (IPTG) (100 µM) was
added when the optical density at 600 nm reached 0.6, and incubation
was continued for 6 h at 37°C. Cells (4.5 g) were harvested by
centrifugation and disrupted with a French press. Chromatography on
DEAE Sepharose was performed as described above for the C. ammoniagenes enzyme with a 0.0 to 0.3 M NaCl gradient.
Dephospho-CoA kinase activity eluted at 0.15 M NaCl. Active fractions
were pooled and loaded onto a Q Sepharose column (1.5 by 10 cm)
equilibrated with Q Sepharose buffer. The column was washed with Q
Sepahrose buffer (30 ml) and eluted with a 150-ml linear KCl gradient
(0.0 to 0.5 M KCl) in the same buffer. Dephospho-CoA kinase activity
eluted at 0.20 M KCl.
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RESULTS |
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Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
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Purification of dephospho-CoA kinase from C. ammoniagenes.
The initial steps for purification of
dephospho-CoA kinase from C. ammoniagenes were performed as
described previously (12). Dephospho-CoA kinase activity
could not be measured in the crude cell lysate due to high background
values but was readily measured in fractions resulting from
chromatography of the crude lysate on DEAE Sepharose (Table
1). Active fractions from the DEAE column were then applied to a Red A agarose column. While most of the protein
was not bound by the column, the dephospho-CoA kinase was bound after a
few minutes of equilibration. The enzyme was selectively eluted by
using a 2 mM solution of ATP, and almost 200-fold purification was
obtained with this step. The enzyme fractions obtained in this way
contained only minor impurities as determined by SDS-PAGE. Final
purification was achieved by further chromatography on Q Sepharose. The
purified enzyme produced one band at a molecular mass of about 25 kDa.
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Cloning, expression, and purification of recombinant dephospho-CoA
kinase from E. coli.
The purified dephospho-CoA kinase
obtained from C. ammoniagenes was subjected to 30 cycles of
N-terminal sequencing, which resulted in the sequence MKIIG LIGGI GSGKT
TVAQL FVDEG FPLVD. This sequence was used to search the nonredundant
protein sequence databases using BLAST (2). The search
revealed a homologous protein (60% identity, 74% positives) encoded
by the yacE gene of E. coli (Fig.
2), as well as homologous proteins (up to
78% identical) from several other sources. The 206-amino-acid protein has a calculated molecular mass of 22,600 Da. The coaE gene
from E. coli W3110 was amplified by PCR, and the resulting
product was inserted into pEt28b(+). The resulting plasmid, designated pEt/coaE, was sequenced by using the T7 promoter primer. E. coli BL21(DE3) containing this plasmid expressed the dephospho-CoA kinase upon induction with IPTG. The recombinant dephospho-CoA kinase
was initially purified by anion-exchange chromatography on DEAE
Sepharose, like the wild-type C. ammoniagenes enzyme (Table 2). The resulting dephospho-CoA kinase
activity was found to not bind to Red A agarose. The DEAE fractions
were then purified further by anion-exchange chromatography on Q
Sepharose. The recombinant dephospho-CoA kinase was found
to be pure by SDS-PAGE. Twelve cycles of N-terminal sequencing of the
recombinant protein gave the sequence ARYIVALTGGIG.
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Preliminary characterization of the recombinant dephospho-CoA kinase. In addition to the standard assay for dephospho-CoA kinase activity based on ADP formation, an assay for CoA formation by the recombinant enzyme was performed by using phosphotransacetylase and acetyl phosphate with monitoring of thioester formation at 240 nm. This assay gave the same activity as the standard assay. A sample resulting from preparative phosphorylation of dephospho-CoA with the recombinant enzyme was subjected to 1H NMR analysis, and the spectrum obtained was identical to that of authentic CoA. The purified enzyme eluted from a calibrated Sephacryl S-100 column with an elution volume equivalent to a molecular mass of 22.2 ± 1.1 kDa, suggesting that the native enzyme exists as a monomer.
Kinetic constants for dephospho-CoA kinase were determined by measuring the initial rates obtained with different concentrations of ATP and dephospho-CoA. Km values of 0.74 and 0.14 mM were determined for dephospho-CoA and ATP, respectively. Activities were also observed when adenosine, APS, and AMP were used as substrates, and the activities observed were 4, 8, and 5% respectively, of the activity when 0.4 mM dephospho-CoA was used as the substrate. The recombinant dephospho-CoA kinase exhibited high activity over a broad pH range, and the maximum activity occurred at about pH 8.5.| |
DISCUSSION |
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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C. ammoniagenes was chosen as a source of dephospho-CoA kinase for purification based on the higher activities of the enzymes of CoA biosynthesis in this bacterium than in E. coli and other microorganisms (18) and because of prior success in our lab with purification of phosphopantetheine adenylyltransferase and partial purification of dephospho-CoA kinase from this organism (12). Purification of the dephospho-CoA kinase turned out to be fairly simple, largely due to the high degree of purification achieved during the very selective elution of dephospho-CoA kinase from Red A agarose when ATP was used. A BLAST search using the N-terminal sequence of the purified enzyme suggested that dephospho-CoA kinase of E. coli was the protein encoded by yacE. The 30 amino acid residues determined for the Corynebacterium dephospho-CoA kinase were 60% identical to the predicted N-terminal amino acid sequence encoded by the E. coli gene (Fig. 2), and the molecular mass of the deduced yacE protein (22.6 kDa) was approximately the same as that of the purified dephospho-CoA kinase (about 25 kDa). While yacE and its homologues have been characterized as hypothetical kinases or ATP-binding proteins, no specific function of the yacE protein has been proposed previously. Cloning and expression of yacE, now renamed coaE, provided the recombinant dephospho-CoA kinase.
The enzyme APS kinase, which catalyzes phosphoryl transfer from ATP to the 3'-hydroxyl group of APS, was previously reported to accept dephospho-CoA as a substrate, although the specific activity was not reported (17). The dephospho-CoA kinase gene does not have significant homology to the APS kinase gene. APS and two other potential alternate substrates, adenosine and AMP, were tested as substrates for the recombinant dephospho-CoA kinase. All three of these compounds showed significant activity, although the activities were much less than that observed when dephospho-CoA was the substrate. These results and the NMR product analysis which showed that dephospho-CoA was phosphorylated on the 3'-hydroxyl group provide evidence that dephospho-CoA is the natural substrate for the recombinant enzyme. We are unaware of any enzymes other than dephospho-CoA kinase and APS kinase that catalyze 3'-phosphorylation of a nucleotide, although DNA and RNA polymerases catalyze 3'-nucleotidylation (24).
The Km determined for dephospho-CoA (0.74 mM) is much higher than the value reported for the dephospho-CoA kinase activity of the partially purified bifunctional enzyme from rat liver (0.01 mM) (19) and somewhat higher than the value previously measured for the C. ammoniagenes dephospho-CoA kinase (0.12 mM) (13). Like the rat liver enzyme, maximum activity is observed at a high pH. The combined subunit molecular weights of the E. coli phosphopantetheine adenylyltransferase (18,000) (7) and dephospho-CoA kinase (22,600) are somewhat less than the molecular weight of the bifunctional enzyme from pork liver (57,000) (25). There is no information concerning the structure of the pork liver enzyme or any other mammalian bifunctional enzyme that permits further analysis of similarities to or differences from the equivalent monofunctional enzymes from E. coli.
Surprisingly, coaE is not clustered with any genes related to CoA biosynthesis or metabolism in any of the microbial genomes sequenced. Notably, some chromosomal clustering is observed between coaE and some genes for NAD de novo biosynthesis. More than 60 homologues of the E. coli coaE gene product were identified by using the WIT system (15), and sequences from a range of organisms are shown in Fig. 2. The different sequences have several areas of high homology, including the Walker kinase motif (22, 23) (residues 9 to 17 of the E. coli enzyme). The homologues in Fig. 2 illustrate that the coaE gene is present in a wide range of organisms, including bacteria, fungi, and animals, although no plant species have been identified. Identification of a human homologue of the monofunctional dephospho-CoA kinase is somewhat surprising, given the evidence that there is a bifunctional phosphopantetheine adenylyltransferase-dephospho-CoA kinase in rat and pork livers (8, 25).
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.
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ACKNOWLEDGMENTS |
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This work was supported by National Institutes of Health grant GM45831. The NMR facility at Stony Brook is supported by National Science Foundation grant CHE9413510. The Center for Analysis and Synthesis of Macromolecules at Stony Brook is supported by National Institutes of Health grant RR02427 and the Center for Biotechnology.
We thank Zuzana Zachar, Stefan Tafrov, Ann Sutton, Rolf Sternglanz, and Peter Tonge for advice and technical assistance. We thank Chi-Huey Wong and coworkers for providing an APS kinase expression plasmid.
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FOOTNOTES |
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* Corresponding author. Mailing address: Department of Chemistry, State University at Stony Brook, Stony Brook, NY 11794-3400. Phone: (631) 632-7923. Fax: (631) 632-7960. E-mail: dale.drueckhammer{at}sunysb.edu.
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REFERENCES |
|---|
|
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Abstract
Introduction
Materials and Methods
Results
Discussion
References
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|---|
| 1. | Abiko, Y. 1975. Metabolism of coenzyme A, p. 1-25. In D. M. Greenberg (ed.), Metabolic pathways, 3rd ed., vol. 7. Academic Press, New York, N.Y. |
| 2. |
Altschul, S. F.,
T. L. Madden,
A. A. Schaffer,
J. Zhang,
Z. Zhang,
W. Miller, and D. J. Lipman.
1997.
Gapped BLAST and PSI-BLAST: a new generation of protein database search programs.
Nucleic Acids Res.
25:3389-3402 |
| 3. | Begley, T. P., C. Kinsland, S. Taylor, M. Tandon, R. Nicewonger, M. Wu, H.-J. Chiu, N. Kelleher, N. Campobasso, and Y. Zhang. 1998. Cofactor biosynthesis: a mechanistic perspective. Top. Curr. Chem. 195:93-142. |
| 4. | Bradford, M. M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72:248-254[CrossRef][Medline]. |
| 5. | Bucovaz, E. T., J. L. Rhoades, J. C. Morrison, J. E. Fryer, W. C. Morrison, and W. D. Whybrew. 1977. Characteristics of coenzyme A-synthesizing protein complex of bakers yeast. Fed. Proc. 36:876. |
| 6. | Bucovaz, E. T., R. M. Macleod, J. C. Morrison, and W. D. Whybrew. 1997. The coenzyme A-synthesizing protein complex and its proposed role in CoA biosynthesis in bakers' yeast. Biochimie 79:787-798[Medline]. |
| 7. |
Geerlof, A.,
A. Lewendon, and W. V. Shaw.
1999.
Purification and characterization of phosphopantetheine adenylyltransferase from Escherichia coli.
J. Biol. Chem.
274:27105-27111 |
| 8. | Hoagland, M. B., and G. D. Novelli. 1953. Biosynthesis of coenzyme A from phosphopantetheine and of pantetheine from pantothenate. J. Biol. Chem. 207:767-773. |
| 9. | Izard, T., and A. Geerlof. 1999. The crystal structure of a novel bacterial adenylyltransferase reveals half of sites reactivity. EMBO J. 18:2021-2030[CrossRef][Medline]. |
| 10. |
Kupke, T.,
M. Uebele,
D. Schmid,
G. Jung,
M. Blaesse, and S. Steinbacher.
2000.
Molecular characterization of lantibiotic-synthesizing enzyme EpiD reveals a function for bacterial Dfp proteins in coenzyme A biosynthesis.
J. Biol. Chem.
275:31838-31846 |
| 11. | Laemmli, U. K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680-685[CrossRef][Medline]. |
| 12. | Martin, D. P., and D. G. Drueckhammer. 1993. Separate enzymes catalyze the final two steps of coenzyme A biosynthesis in Brevibacterium ammoniagenes: purification of pantetheine phosphate adenylyltransferase. Biochem. Biophys. Res. Commun. 192:1155-1161[CrossRef][Medline]. |
| 13. | Martin, D. P., R. T. Bibart, and D. G. Drueckhammer. 1994. Synthesis of novel analogs of acetyl coenzyme A: mimics of enzyme reaction intermediates. J. Am. Chem. Soc. 116:4660-4668[CrossRef]. |
| 14. | Merkel, W. K., and B. P. Nichols. 1996. Characterization and sequence of the Escherichia coli panBCD gene cluster. FEMS Microbiol. Lett. 143:247-252[CrossRef][Medline]. |
| 15. |
Overbeek, R.,
N. Larsen,
G. D. Pusch,
M. Dsouza,
E. Selkov, Jr.,
N. Kyrpides,
M. Fonstein,
N. Maltsev, and E. Selkov.
2000.
WIT: integrated system for high-throughput genome sequence analysis and metabolic reconstruction.
Nucleic Acids Res.
28:123-125 |
| 16. | Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. |
| 17. | Satishchandran, C., J. C. Taylor, and G. D. Markham. 1993. The ORF1 of the gentamicin-resistance operon (aac) of Pseudomonas aeruginosa encodes adenosine 5-phosphosulfate kinase. Mol. Microbiol. 9:1223-1227[CrossRef][Medline]. |
| 18. | Shimizu, S., K. Kubo, H. Morioka, Y. Tani, and K. Ogata. 1974. Studies on metabolism of pantothenic acid in microorganisms. 9. Some aspects of enzyme activities involved in coenzyme A biosyntheis in various microorganisms. Agric. Biol. Chem. 38:1015-1021. |
| 19. | Skrede, S., and O. Halvorsen. 1983. Mitochondrial pantetheinephosphate adenylyltransferase and dephospho-CoA kinase. Eur. J. Biochem. 131:57-63[Medline]. |
| 20. |
Song, W. J., and S. Jackowski.
1992.
Cloning, sequencing, and expression of the pantothenate kinase (coaA) gene of Escherichia coli.
J. Bacteriol.
174:6411-6417 |
| 21. |
Thompson, J. D.,
D. G. Higins, and T. J. Gibson.
1994.
CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting positions-specific gap penalties and weight matrix choice.
Nucleic Acids Res.
22:4673-4680 |
| 22. | Traut, T. W. 1994. The functions and consensus motifs of nine types of peptide segments that form different types of nucleotide-binding sites. Eur. J. Biochem. 222:9-19[Medline]. |
| 23. |
Walker, J. E.,
M. Saraste,
M. J. Runswick, and N. J. Gay.
1982.
Distantly related sequences in the  - and -subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold.
EMBO J.
1:945-951[Medline].
|
| 24. | Webb, E. C. 1992. Enzyme nomenclature. Academic Press, San Diego, Calif. |
| 25. | Worrall, D. M., and P. K. Tubbs. 1983. A bifunctional enzyme complex in coenzyme A biosynthesis: purification of pantetheine phosphate adenylyltransferase and dephospho-CoA kinase. Biochem. J. 215:153-157[Medline]. |
| 26. | Zheng, R., and J. S. Blanchard. 2000. Kinetic and mechanistic analysis of the E. coli panE-encoded ketopantoate reductase. Biochemistry 39:3708-3717[CrossRef][Medline]. |
Journal of Bacteriology, May 2001, p. 2774-2778, Vol. 183, No. 9
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.9.2774-2778.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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Choudhry, A. E., Mandichak, T. L., Broskey, J. P., Egolf, R. W., Kinsland, C., Begley, T. P., Seefeld, M. A., Ku, T. W., Brown, J. R., Zalacain, M., Ratnam, K.
(2003). Inhibitors of Pantothenate Kinase: Novel Antibiotics for Staphylococcal Infections. Antimicrob. Agents Chemother.
47: 2051-2055
[Abstract] [Full Text] -
Rock, C. O., Park, H.-W., Jackowski, S.
(2003). Role of Feedback Regulation of Pantothenate Kinase (CoaA) in Control of Coenzyme A Levels in Escherichia coli. J. Bacteriol.
185: 3410-3415
[Abstract] [Full Text] -
Strauss, E., Begley, T. P.
(2002). The Antibiotic Activity of N-Pentylpantothenamide Results from Its Conversion to Ethyldethia-Coenzyme A, a Coenzyme A Antimetabolite. J. Biol. Chem.
277: 48205-48209
[Abstract] [Full Text] -
Kupke, T.
(2002). Molecular Characterization of the 4'-Phosphopantothenoylcysteine Synthetase Domain of Bacterial Dfp Flavoproteins. J. Biol. Chem.
277: 36137-36145
[Abstract] [Full Text] -
Gerdes, S. Y., Scholle, M. D., D'Souza, M., Bernal, A., Baev, M. V., Farrell, M., Kurnasov, O. V., Daugherty, M. D., Mseeh, F., Polanuyer, B. M., Campbell, J. W., Anantha, S., Shatalin, K. Y., Chowdhury, S. A. K., Fonstein, M. Y., Osterman, A. L.
(2002). From Genetic Footprinting to Antimicrobial Drug Targets: Examples in Cofactor Biosynthetic Pathways. J. Bacteriol.
184: 4555-4572
[Abstract] [Full Text] -
Daugherty, M., Polanuyer, B., Farrell, M., Scholle, M., Lykidis, A., de Crecy-Lagard, V., Osterman, A.
(2002). Complete Reconstitution of the Human Coenzyme A Biosynthetic Pathway via Comparative Genomics. J. Biol. Chem.
277: 21431-21439
[Abstract] [Full Text] -
Rubio, A., Downs, D. M.
(2002). Elevated Levels of Ketopantoate Hydroxymethyltransferase (PanB) Lead to a Physiologically Significant Coenzyme A Elevation in Salmonella enterica Serovar Typhimurium. J. Bacteriol.
184: 2827-2832
[Abstract] [Full Text] -
Kupke, T.
(2001). Molecular Characterization of the 4'-Phosphopantothenoylcysteine Decarboxylase Domain of Bacterial Dfp Flavoproteins. J. Biol. Chem.
276: 27597-27604
[Abstract] [Full Text]
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