Crystal structure of UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase from Escherichia coli

Bertrand, J. A.; Auger, Geneviève; Fanchon, Éric; Martin, Lydie; Blanot, Didier; Heijenoort, Jean van; Dideberg, Otto

doi:10.1093/emboj/16.12.3416

Cited by 132 publications

(97 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1) shows sequence similarity to members of a superfamily of peptide ligases [30,31] that includes certain murein ligases and folyl poly-gglutamate ligase. Members of this superfamily contain an ATP-binding P-loop motif, and show a Rossman fold [32]. Further residues that have been found to be invariant in some of those ligases are conserved in cyanophycin synthetase [16].…”

Section: R E S U L T S a N D Discussionmentioning

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

Section: R E S U L T S a N D Discussionmentioning

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

show abstract

“…Crystals of conformationally closed MurD ligase were obtained by co-crystallising MurD at 293 K with UMA and a nonhydrolysable analogue of ATP (AMP-PNP) [11,12,32]. The crystals were obtained using the vapour-diffusion method and a hangingdrop system in Linbro plates, by mixing drops of 2 mL of MurD solution (3 mg/mL purified enzyme, 20 mM HEPES, pH 7.4, 200 mM NaCl, 5 mM dithiothreitol, 1 mM UMA and 5 mM AMP-PNP) and 2 mL of reservoir solution (0.1 M HEPES, pH 7.5, 1.9 M ammonium sulphate, 7% (w/v) PEG 400 and 50 mM NaCl).…”

Section: Crystallisation Preparation Of the Inhibitor Complex And Dmentioning

confidence: 99%

Second-generation sulfonamide inhibitors of d-glutamic acid-adding enzyme: Activity optimisation with conformationally rigid analogues of d-glutamic acid

Sosič¹,

Barreteau²,

Simčič³

et al. 2011

European Journal of Medicinal Chemistry

View full text Add to dashboard Cite

a b s t r a c t D-Glutamic acid-adding enzyme (MurD) catalyses the essential addition of D-glutamic acid to the cytoplasmic peptidoglycan precursor UDP-N-acetylmuramoyl-L-alanine, and as such it represents an important antibacterial drug-discovery target enzyme. Based on a series of naphthalene-N-sulfonyl-DGlu derivatives synthesised recently, we synthesised two series of new, optimised sulfonamide inhibitors of MurD that incorporate rigidified mimetics of D-Glu. The compounds that contained either constrained D-Glu or related rigid D-Glu mimetics showed significantly better inhibitory activities than the parent compounds, thereby confirming the advantage of molecular rigidisation in the design of MurD inhibitors. The binding modes of the best inhibitors were examined with high-resolution NMR spectroscopy and X-ray crystallography. We have solved a new crystal structure of the complex of MurD with an inhibitor bearing a 4-aminocyclohexane-1,3-dicarboxyl moiety. These data provide an additional step towards the development of sulfonamide inhibitors with potential antibacterial activities.

show abstract

“…In recent years, intense effort has been focused on determining the structures of the enzymes that synthesize peptidoglycan. Structures of several of the early enzymes in the biosynthetic pathway have been reported~Fan et al, 1994;Benson et al, 1995;Skarzynski et al, 1996;Bertrand et al, 1997! ; however, the later enzymes have proven more difficult to study because both they and their substrates are membrane-associated.…”

mentioning

confidence: 99%

The 1.9 Å crystal structure of Escherichia coli MurG, a membrane‐associated glycosyltransferase involved in peptidoglycan biosynthesis

et al. 2000

View full text Add to dashboard Cite

The 1.9 Å X-ray structure of a membrane-associated glycosyltransferase involved in peptidoglycan biosynthesis is reported. This enzyme, MurG, contains two a0b open sheet domains separated by a deep cleft. Structural analysis suggests that the C-terminal domain contains the UDP-GlcNAc binding site while the N-terminal domain contains the acceptor binding site and likely membrane association site. Combined with sequence data from other MurG homologs, this structure provides insight into the residues that are important in substrate binding and catalysis. We have also noted that a conserved region found in many UDP-sugar transferases maps to a b0a0b0a supersecondary structural motif in the donor binding region of MurG, an observation that may be helpful in glycosyltransferase structure prediction. The identification of a conserved structural motif involved in donor binding in different UDP-sugar transferases also suggests that it may be possible to identify-and perhaps alter-the residues that help determine donor specificity.

show abstract

Crystal structure of UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase from Escherichia coli

Cited by 132 publications

References 53 publications

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

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

Second-generation sulfonamide inhibitors of d-glutamic acid-adding enzyme: Activity optimisation with conformationally rigid analogues of d-glutamic acid

The 1.9 Å crystal structure of Escherichia coli MurG, a membrane‐associated glycosyltransferase involved in peptidoglycan biosynthesis

Contact Info

Product

Resources

About