Structure of Human Phosphopantothenoylcysteine Synthetase at 2.3 Å Resolution

Manoj, Narayanan; Strauss, Erick; Begley, Tadhg P.; Ealick, S.E.

doi:10.1016/s0969-2126(03)00146-1

Cited by 31 publications

(36 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Identical residues are in red. ( b ) The location of mutation sites in the three-dimensional structure of human PPCS [40]. T48I mutation site in ppc1-537 locates near the catalytic centre, while the site for ppc1-88 locates at the periphery of PPCS.…”

Section: Resultsmentioning

confidence: 99%

“…It is similar to human PPCS (amino acid identity 42%) and budding yeast Cab2 (identity 45%). Two mutation sites in the three-dimensional structure of PPCS/Ppc1 based on the human PPCS [40] are shown in figure 3 b . The mutation site T48I in ppc1-537 resides closely to the central catalytic domain, whereas the M209T site in ppc1-88 is located at the periphery of molecule.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Impaired coenzyme A synthesis in fission yeast causes defective mitosis, quiescence-exit failure, histone hypoacetylation and fragile DNA

et al. 2012

View full text Add to dashboard Cite

Biosynthesis of coenzyme A (CoA) requires a five-step process using pantothenate and cysteine in the fission yeast Schizosaccharomyces pombe. CoA contains a thiol (SH) group, which reacts with carboxylic acid to form thioesters, giving rise to acyl-activated CoAs such as acetyl-CoA. Acetyl-CoA is essential for energy metabolism and protein acetylation, and, in higher eukaryotes, for the production of neurotransmitters. We isolated a novel S. pombe temperature-sensitive strain ppc1-537 mutated in the catalytic region of phosphopantothenoylcysteine synthetase (designated Ppc1), which is essential for CoA synthesis. The mutant becomes auxotrophic to pantothenate at permissive temperature, displaying greatly decreased levels of CoA, acetyl-CoA and histone acetylation. Moreover, ppc1-537 mutant cells failed to restore proliferation from quiescence. Ppc1 is thus the product of a super-housekeeping gene. The ppc1-537 mutant showed combined synthetic lethal defects with five of six histone deacetylase mutants, whereas sir2 deletion exceptionally rescued the ppc1-537 phenotype. In synchronous cultures, ppc1-537 cells can proceed to the S phase, but lose viability during mitosis failing in sister centromere/kinetochore segregation and nuclear division. Additionally, double-strand break repair is defective in the ppc1-537 mutant, producing fragile broken DNA, probably owing to diminished histone acetylation. The CoA-supported metabolism thus controls the state of chromosome DNA.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Impaired coenzyme A synthesis in fission yeast causes defective mitosis, quiescence-exit failure, histone hypoacetylation and fragile DNA

et al. 2012

View full text Add to dashboard Cite

show abstract

“…Although the observed molecular mass deviates by 46.8 kDa from the theoretical value for a homododecamer, we assume (taking the inaccuracy of molecular weight determinations by gel filtration into account) that His-CoaC builds up homododecamers. In conclusion, the M. jannaschii Dfp protein is a also a homododecameric enzyme, with the CoaC domain forming a dodecameric core structure as has recently been modeled for the E. coli enzyme (20). bifunctional Dfp protein into CoaB and CoaC.…”

Section: Resultsmentioning

confidence: 85%

4′-Phosphopantetheine Biosynthesis in Archaea

Kupke

Schwarz²

2006

Journal of Biological Chemistry

View full text Add to dashboard Cite

Coenzyme A (CoA) and 2 are essential cofactors for many enzymatic reactions, and acyl-CoA derivatives are key intermediates in energy metabolism. 4Ј-Phosphopantetheine is a cofactor of enzymes that play a role in the biosynthesis of fatty acids, polypeptide antibiotics, and polyketides. The thiol group of the cysteamine moiety of coenzyme A is the functional group because it activates substrates as thioesters.Coenzyme A is synthesized from pantothenate in five steps, and all of the genes involved in the biosynthesis of the cofactor in eubacteria, plants, and human have recently been cloned and characterized. In the first step, pantothenate is phosphorylated to 4Ј-phosphopantothenate by pantothenate kinase, which is encoded by the coaA gene. Then, (R)-4Ј-phospho-N-pantothenoylcysteine (PPC) is synthesized by the addition of cysteine to 4Ј-phosphopantothenate (CoaB activity), and in the next step, PPC is decarboxylated to 4Ј-phosphopantetheine (CoaC activity). 4Ј-Phosphopantetheine is converted to coenzyme A by the enzymes 4Ј-phosphopantetheine adenylyltransferase (CoaD) and dephospho-CoA kinase (CoaE; reviewed in Ref. 1).The key reaction in coenzyme A biosynthesis, the synthesis of the phosphopeptide-like cofactor 4Ј-phosphopantetheine from 4Ј-phosphopantothenate and cysteine, is catalyzed in Escherichia coli and in most eubacteria by the bifunctional Dfp (CoaBC) flavoprotein in a multistep process (Fig. 1) (2). 4Ј-Phosphopantothenate is activated by reaction with CTP; the 4Ј-phosphopantothenoyl-cytidylate formed is attacked by cysteine, and PPC is synthesized (2-4). Crystal structure analysis of E. coli CoaB (5) revealed the 4Ј-phosphopantothenate and CTP binding motifs of CoaB. The nucleobase binding motif II of eubacterial CTP-binding CoaBs deviates from that of eukaryotic ATP-binding PPC synthetases. In E. coli Dfp the sequence reads 307 NPDIV, whereas the sequence is 210 VPKLL in the human enzyme (residues shown in boldface are those conserved in eubacterial/eukaryotic enzymes). In the next step, PPC is oxidatively decarboxylated to 4Ј-phosphopantothenoylaminoenethiol by the NH 2 -terminal FMNbinding CoaC domain of Dfp (6 -9). Subsequent reduction of 4Ј-phosphopantothenoylaminoenethiol to 4Ј-phosphopantetheine by the reduced cofactor FMNH 2 depends on the conserved cysteine residue of the 16-amino acid 4Ј-phosphopantothenoylcysteine binding clamp, 151 PDSGSQACGDIGPGRM (6,8,9). Binding of 4Ј-phosphopantothenoylcysteine also involves the Asn residue of the PXMNXXMW motif, which contacts the carboxyl group (8, 10). The presence of conserved CoA biosynthetic genes indicates that all Archaea convert 4Ј-phosphopantothenate into coenzyme A by using CoaB, CoaC, CoaD, and CoaE activities, although cloning and functional characterization of archaebacterial coenzyme A biosynthetic genes has not been published until now (11). In this paper, we have characterized the bifunctional Dfp protein of Methanocaldococcus jannaschii. The 5Ј-coaC part and the 3Ј-coaB part of the archaebacterial dfp gene were expressed in E. coli,...

show abstract

“…This is in accordance with the previous finding that PPCS catalyzes the conversion of pantothenate to coenzyme A by utilizing ATP as a substrate. 23 In another example, K375 in protein disulfide-isomerase (PDI) exhibits low R ATP10/1 ratios in ATP profiling experiments for both cell lines, strongly suggesting that this protein possesses ATP-binding property. PDI is a multifunction protein and it is involved in protein folding.…”

Section: Resultsmentioning

confidence: 99%

Isotope-Coded ATP Probe for Quantitative Affinity Profiling of ATP-Binding Proteins

2013

View full text Add to dashboard Cite

ATP-binding proteins play significant roles in numerous cellular processes. Here, we introduced a novel isotope-coded ATP-affinity probe (ICAP) as acylating agent to simultaneously enrich and incorporate isotope label to ATP-binding proteins. By taking advantage of the quantitative capability of this isotope-coded probe, we devised an affinity profiling strategy to comprehensively characterize ATP-protein interactions at the entire proteome scale. False-positive identification of ATP-binding sites derived from non-specific labeling was effectively minimized through the comparison of the labeling behaviors of lysine residues with the use of low and high concentrations of the ICAP reagents. A total of 258 previously known ATP-binding proteins from lysates of Hela-S3 and Jurkat-T cells were validated with this affinity profiling assay. Additionally, we demonstrated that this novel quantitative ATP-affinity profiling strategy is particularly useful for unveiling previously unrecognized nucleotide-binding sites in ATP-binding proteins. For example, our profiling results revealed K356 as a new ATP-binding site in HSP90. Furthermore, 293 proteins without documented ATP-binding GO were predicted to be ATP-binding proteins on the basis of our quantitative affinity profiling results. We also uncovered, for the first time, the ATP-binding capability of human proliferating cell nuclear antigen (PCNA), identified the lysine residue involved in ATP binding, and validated the protein’s capacity in ATP binding with an independent assay. The ICAP approach described in the present paper should be generally applicable for the quantitative assessment of ATP-binding proteins in proteomic samples from cells and tissues.

show abstract

Structure of Human Phosphopantothenoylcysteine Synthetase at 2.3 Å Resolution

Cited by 31 publications

References 35 publications

Impaired coenzyme A synthesis in fission yeast causes defective mitosis, quiescence-exit failure, histone hypoacetylation and fragile DNA

Impaired coenzyme A synthesis in fission yeast causes defective mitosis, quiescence-exit failure, histone hypoacetylation and fragile DNA

4′-Phosphopantetheine Biosynthesis in Archaea

Isotope-Coded ATP Probe for Quantitative Affinity Profiling of ATP-Binding Proteins

Contact Info

Product

Resources

About