Formation of adenosine 5′‐tetraphosphate from the acyl phosphate intermediate: a difference between the MurC and MurD synthetases of <i>Escherichia coli</i>

Bouhss, Ahmed; Dementin, Sébastien; Heijenoort, Jean van; Parquet, Claudine; Blanot, Didier

doi:10.1016/s0014-5793(99)00684-5

Cited by 27 publications

(18 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…As established with other Mur ligases, the MurE reaction presumably proceeds by phosphorylation of the C-terminal carboxylate of UMAG by the ␥-phosphate of ATP to form an acyl phosphate intermediate, followed by nucleophilic attack by the ␣-amino group of A 2 pm to produce UMT ( Fig. 1), ADP, and inorganic phosphate (13)(14)(15). This mechanism is supported by the three-dimensional structures of MurD and MurD complexes.…”

supporting

confidence: 58%

Crystal Structure of UDP-N-acetylmuramoyl-l-alanyl-d-glutamate:meso-Diaminopimelate Ligase from Escherichia Coli

Gordón

Flouret

Chantalat

et al. 2001

Journal of Biological Chemistry

Self Cite

136

134

View full text Add to dashboard Cite

UDP-N-acetylmuramoyl-L-alanyl-D-glutamate:meso-diaminopimelate ligase is a cytoplasmic enzyme that catalyzes the addition of meso-diaminopimelic acid to nucleotide precursor UDP-N-acetylmuramoyl-L-alanyl-D-glutamate in the biosynthesis of bacterial cell-wall peptidoglycan. The crystal structure of the Escherichia coli enzyme in the presence of the final product of the enzymatic reaction, UDP-MurNAc-L-Ala-␥-D-Glu-meso-A 2 pm, has been solved to 2.0 Å resolution. Phase information was obtained by multiwavelength anomalous dispersion using the K shell edge of selenium. The protein consists of three domains, two of which have a topology reminiscent of the equivalent domain found in the already established three-dimensional structure of the UDP-N-acetylmuramoyl-L-alanine: D-glutamateligase (MurD) ligase, which catalyzes the immediate previous step of incorporation of D-glutamic acid in the biosynthesis of the peptidoglycan precursor. The refined model reveals the binding site for UDP-MurNAc-LAla-␥-D-Glu-meso-A 2 pm, and comparison with the six known MurD structures allowed the identification of residues involved in the enzymatic mechanism. Interestingly, during refinement, an excess of electron density was observed, leading to the conclusion that, as in MurD, a carbamylated lysine residue is present in the active site. In addition, the structural determinant responsible for the selection of the amino acid to be added to the nucleotide precursor was identified.Peptidoglycan, the polymeric mesh of the bacterial cell wall, plays a critical role in protecting bacteria against osmotic lysis. It consists of linear repeating disaccharide chains cross-linked by short peptide bridges. During the cytoplasmic steps involved in the biosynthesis of the peptidoglycan precursor, four ADPforming ligases (namely the Mur ligases) catalyze the assembly of the peptide moiety by the successive addition of L-alanine, D-glutamate, diaminopimelic acid, or L-lysine, and, finally, dipeptide D-alanyl-D-alanine to UDP-N-acetylmuramic acid (1, 2). Because all these enzymes are essential for cell viability, they are attractive targets for antibacterial chemotherapy. In Escherichia coli, these ligases are the products of the murC, murD, murE, and murF genes, located in the mra region (3). Sequence comparison of the four E. coli Mur ligases shows several homologous regions, suggesting that these enzymes may be evolutionarily related and may use similar enzymatic mechanisms (4 -6). In earlier publications, we reported the structure of UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase (MurD), 1 both in the native form and complexed with substrates (7-9). MurD consists of three domains with topologies reminiscent of a nucleotide-binding fold; the N-and Cterminal domains have a dinucleotide-binding fold (the Rossmann fold), and the central domain displays a mononucleotidebinding fold, also seen in ATP-binding proteins. A comparison of six MurD structures reveals that large C-terminal rotation, loop rearrangement, and subdomain movements occur upon subs...

show abstract

supporting

confidence: 58%

Crystal Structure of UDP-N-acetylmuramoyl-l-alanyl-d-glutamate:meso-Diaminopimelate Ligase from Escherichia Coli

Gordón

Flouret

Chantalat

et al. 2001

Journal of Biological Chemistry

Self Cite

136

134

View full text Add to dashboard Cite

show abstract

“…3, gray shaded region), the Cys-S P O 2 -moiety (Cys52 in human PrxI, hPrxI) is phosphorylated by the g-phosphate of ATP to form the sulfinic phosphoryl ester (Cys-S P O 2 PO 3 2-) (6). This type of ATP-mediated activation is reminiscent of the activation of carboxyl groups in a variety of biologic processes, but is novel for sulfur chemistry (7,16,17). A thiosulfinate intermediate (Prx-S P O-S-Srx) is then formed, after the attack of a conserved Cys residue in Srx (Cys 99 in hSrx).…”

Section: Molecular Basis For Srx Actionmentioning

confidence: 99%

Reduction of Cysteine Sulfinic Acid in Eukaryotic, Typical 2-Cys Peroxiredoxins by Sulfiredoxin

Lowther

Haynes

2011

Antioxidants & Redox Signaling

View full text Add to dashboard Cite

The eukaryotic, typical 2-Cys peroxiredoxins (Prxs) are inactivated by hyperoxidation of one of their active-site cysteine residues to cysteine sulfinic acid. This covalent modification is thought to enable hydrogen peroxidemediated cell signaling and to act as a functional switch between a peroxidase and a high-molecular-weight chaperone. Moreover, hyperoxidation has been implicated in a variety of disease states associated with oxidative stress, including cancer and aging-associated pathologies. A repair enzyme, sulfiredoxin (Srx), reduces the sulfinic acid moiety by using an unusual ATP-dependent mechanism. In this process, the Prx molecule undergoes dramatic structural rearrangements to facilitate repair. Structural, kinetic, mutational, and mass spectrometry-based approaches have been used to dissect the molecular basis for Srx catalysis. The available data support the direct formation of Cys sulfinic acid phosphoryl ester and protein-based thiosulfinate intermediates. This review discusses the role of Srx in the reversal of Prx hyperoxidation, the questions raised concerning the reductant required for human Srx regeneration, and the deglutathionylating activity of Srx. The complex interplay between Prx hyperoxidation, other forms of Prx covalent modification, and the oligomeric state also are discussed. Antioxid. Redox Signal. 15, 99-109.

show abstract

“…The energy for peptide bond formation is provided by ATP which is hydrolysed to ADP and P i [16], suggesting that either the amino-acid substrates or the cyanophycin primer are activated by phosphorylation of carboxyl groups. Acylphosphate intermediates are known to be involved in the synthesis of other nonribosomally made peptides such as glutathione [17], poly(g-d-glutamate) [18] and the peptide cross-bridges of murein [19,20].…”

mentioning

confidence: 99%

Biosynthesis of the cyanobacterial reserve polymer multi‐L‐arginyl‐poly‐L‐aspartic acid (cyanophycin)

Berg

Ziegler

Piotukh

et al. 2000

European Journal of Biochemistry

118

100

View full text Add to dashboard Cite

Biosynthesis of the cyanobacterial nitrogen reserve cyanophycin (multi-l-arginyl-poly-l-aspartic acid) is catalysed by cyanophycin synthetase, an enzyme that consists of a single kind of polypeptide. Efficient synthesis of the polymer requires ATP, the constituent amino acids aspartic acid and arginine, and a primer like cyanophycin. Using synthetic peptide primers, the course of the biosynthetic reaction was studied. The following results were obtained: (a) sequence analysis suggests that cyanophycin synthetase has two ATP-binding sites and hence probably two active sites; (b) the enzyme catalyses the formation of cyanophycin-like polymers of 25±30 kDa apparent molecular mass in vitro; (c) primers are elongated at their C-terminus; (d) the constituent amino acids are incorporated stepwise, in the order aspartic acid followed by arginine, into the growing polymer. A mechanism for the cyanophycin synthetase reaction is proposed; (e) the specificity of the enzyme for its amino-acid substrates was also studied. Glutamic acid cannot replace aspartic acid as the acidic amino acid, whereas lysine can replace arginine but is incorporated into cyanophycin at a much lower rate.

show abstract

Formation of adenosine 5′‐tetraphosphate from the acyl phosphate intermediate: a difference between the MurC and MurD synthetases of Escherichia coli

Cited by 27 publications

References 28 publications

Crystal Structure of UDP-N-acetylmuramoyl-l-alanyl-d-glutamate:meso-Diaminopimelate Ligase from Escherichia Coli

Crystal Structure of UDP-N-acetylmuramoyl-l-alanyl-d-glutamate:meso-Diaminopimelate Ligase from Escherichia Coli

Reduction of Cysteine Sulfinic Acid in Eukaryotic, Typical 2-Cys Peroxiredoxins by Sulfiredoxin

Biosynthesis of the cyanobacterial reserve polymer multi‐L‐arginyl‐poly‐L‐aspartic acid (cyanophycin)

Contact Info

Product

Resources

About