GE2270A-resistant Mutations in Elongation Factor Tu Allow Productive Aminoacyl-tRNA Binding to EF-Tu·GTP·GE2270A Complexes

Zuurmond, Anne‐Marie; Graaf, J.Martien de; Olsthoorn-Tieleman, Lian N.; Duyl, Bianca Y. van; Mörhle, Volker G.; Jurnak, Frances; Mesters, J.R.; Hilgenfeld, Rolf; Kraal, Barend

doi:10.1006/jmbi.2000.4260

Cited by 21 publications

(15 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Furthermore, selection on inhibitory concentrations of LFF571 resulted in a substitution at the C. difficile residue analogous to G257 in E. coli EF-Tu. Substitutions at E. coli G257 render the protein resistant to the inhibitory activity of GE2270 A when measured in an in vitro translation assay (23). Along with our previous observation that LFF571 interacts directly with E. coli EF-Tu in vitro (9), these data support the hypothesis that LFF571 inhibits C. difficile translation by binding EF-Tu.…”

Section: ϫ11supporting

confidence: 81%

Mechanism of Action of and Mechanism of Reduced Susceptibility to the Novel Anti-Clostridium difficile Compound LFF571

Leeds

Sachdeva

Mullin

et al. 2012

Antimicrob Agents Chemother

View full text Add to dashboard Cite

bLFF571 is a novel semisynthetic thiopeptide and potent inhibitor of Gram-positive bacteria. We report that the antibacterial activity of LFF571 against Clostridium difficile is due to inhibition of translation. Single-step mutants of C. difficile with reduced susceptibility to LFF571 were selected at frequencies of <4.5 ؋ 10 ؊11 to 1.2 ؋ 10 ؊9 . Sequencing revealed a G260E substitution in the thiopeptide-binding pocket of elongation factor Tu. Importantly, this mutation did not confer cross-resistance to clinically used antimicrobials. These results support the development of LFF571 as a treatment for C. difficile infection.

show abstract

Section: ϫ11supporting

confidence: 81%

Mechanism of Action of and Mechanism of Reduced Susceptibility to the Novel Anti-Clostridium difficile Compound LFF571

Leeds

Sachdeva

Mullin

et al. 2012

Antimicrob Agents Chemother

View full text Add to dashboard Cite

show abstract

“…Both GE37468 and GE2270 were isolated in screens based on the ability of pure EF-Tu to antagonize their antibiotic activity against S. aureus and Moraxella caviae (20). They show similar antibiotic profiles, but notably differ in their inhibition of Streptococcus species (21). They share a triazole-substituted pyridine core and similar primary amino acid sequence with thiomuracin, but GE37468 harbors several distinct modifications in the macrocycle (SI Appendix, Fig.…”

mentioning

confidence: 99%

Identification of the thiazolyl peptide GE37468 gene cluster from Streptomyces ATCC 55365 and heterologous expression in Streptomyces lividans

Young

Walsh

2011

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Thiazolyl peptides are bacterial secondary metabolites that potently inhibit protein synthesis in Gram-positive bacteria and malarial parasites. Recently, our laboratory and others reported that this class of trithiazolyl pyridine-containing natural products is derived from ribosomally synthesized preproteins that undergo a cascade of posttranslational modifications to produce architecturally complex macrocyclic scaffolds. Here, we report the gene cluster responsible for production of the elongation factor Tu (EF-Tu)-targeting 29-member thiazolyl peptide GE37468 from Streptomyces ATCC 55365 and its heterologous expression in the model host Streptomyces lividans . GE37468 harbors an unusual β -methyl- δ -hydroxy-proline residue that may increase conformational rigidity of the macrocycle and impart reduced entropic costs of target binding. Isotope feeding and gene knockout were employed in the engineered S. lividans strain to identify the P450 monooxygenase GetJ as the enzyme involved in posttranslational transformation of isoleucine 8 to β -methyl- δ -hydroxy-proline through a predicted tandem double hydroxylation/cyclization mechanism. Loss of Ile8 oxygenative cyclization or mutation of Ile8 to alanine via preprotein gene replacement resulted in a 4-fold and 2-fold drop in antibiotic activity, respectively. This report of genetic manipulation of a 29-member thiazolyl peptide sets the stage for further genetic examination of structure activity relationships in the EF-Tu targeting class of thiazolyl peptides.

show abstract

“…All mutants expressed EF-Tu with substitutions in Gly257 or Gly275 (E. coli residue numbering). These residues are homologous to residues G257 and G275 in E. coli EF-Tu, which reside in the thiopeptide binding pocket of the enzyme and in which amino acid changes were identified in GE2270 A-resistant E. coli EF-Tu (21,25). Two tuf genes are found in E. faecium, but changes associated with decreased susceptibility were limited to the tufA gene.…”

Section: Resultsmentioning

confidence: 96%

In Vitro and In Vivo Activities of Novel, Semisynthetic Thiopeptide Inhibitors of Bacterial Elongation Factor Tu

Leeds

LaMarche²,

Brewer³

et al. 2011

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Recently, we identified aminothiazole derivatives of GE2270 A. These novel semisynthetic congeners, like GE2270 A, target the essential bacterial protein elongation factor Tu (EF-Tu). Medicinal chemistry optimization of lead molecules led to the identification of preclinical development candidates 1 and 2. These cycloalklycarboxylic acid derivatives show activity against difficult to treat Gram-positive pathogens and demonstrate increased aqueous solubility compared to GE2270 A. We describe here the in vitro and in vivo activities of compounds 1 and 2 compared to marketed antibiotics. Compounds 1 and 2 were potent against clinical isolates of methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci (MIC 90 < 0.25 g/ml) but weaker against the streptococci (MIC 90 > 4 g/ml).

show abstract

GE2270A-resistant Mutations in Elongation Factor Tu Allow Productive Aminoacyl-tRNA Binding to EF-Tu·GTP·GE2270A Complexes

Cited by 21 publications

References 22 publications

Mechanism of Action of and Mechanism of Reduced Susceptibility to the Novel Anti-Clostridium difficile Compound LFF571

Mechanism of Action of and Mechanism of Reduced Susceptibility to the Novel Anti-Clostridium difficile Compound LFF571

Identification of the thiazolyl peptide GE37468 gene cluster from Streptomyces ATCC 55365 and heterologous expression in Streptomyces lividans

In Vitro and In Vivo Activities of Novel, Semisynthetic Thiopeptide Inhibitors of Bacterial Elongation Factor Tu

Contact Info

Product

Resources

About