Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim

@article{Heaslet2009StructuralCO,
  title={Structural comparison of chromosomal and exogenous dihydrofolate reductase from Staphylococcus aureus in complex with the potent inhibitor trimethoprim},
  author={Holly Heaslet and Melissa S. Harris and Kelly C. Fahnoe and Ronald W. Sarver and Henry Putz and Jeanne S. Chang and Chakrapani Subramanyam and G Barreiro and J. Richard Miller},
  journal={Proteins: Structure},
  year={2009},
  volume={76}
}
  • H. Heaslet, M. Harris, +6 authors J. Richard Miller
  • Published 15 August 2009
  • Biology, Medicine
  • Proteins: Structure
Dihydrofolate reductase (DHFR) is the enzyme responsible for the NADPH‐dependent reduction of 5,6‐dihydrofolate to 5,6,7,8‐tetrahydrofolate, an essential cofactor in the synthesis of purines, thymidylate, methionine, and other key metabolites. Because of its importance in multiple cellular functions, DHFR has been the subject of much research targeting the enzyme with anticancer, antibacterial, and antimicrobial agents. Clinically used compounds targeting DHFR include methotrexate for the… 
Increased substrate affinity in the Escherichia coli L28R dihydrofolate reductase mutant causes trimethoprim resistance.
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The experimental and computational analyses together reveal the changes that occur in the energetic landscape of DHFR upon the resistance-conferring L28R mutation, and lays the framework to study structural changes in other trimethoprim resistant DHFR mutants.
Inhibition of Antibiotic-Resistant Staphylococcus aureus by the Broad-Spectrum Dihydrofolate Reductase Inhibitor RAB1
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RAB1 was most effective against Staphylococcus aureus, including methicillin- and vancomycin-resistant (MRSA/VRSA) strains and found broad-spectrum applicability, particularly with regard to Gram-positive organisms.
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Towards the understanding of resistance mechanisms in clinically isolated trimethoprim-resistant, methicillin-resistant Staphylococcus aureus dihydrofolate reductase.
TLDR
A crystal structure of a complex of this double mutant with a lead propargyl-linked antifolate suggests a resistance mechanism consistent both for the propargy-linked class of antifolates and for trimethoprim that is based on the loss of a conserved water-mediated hydrogen bond.
Trimethoprim and other nonclassical antifolates an excellent template for searching modifications of dihydrofolate reductase enzyme inhibitors
TLDR
An excellent review of DHFR inhibition and their relevance to antimicrobial and parasitic chemotherapy was presented and advances in design, synthesis, and biological evaluations in structural modifications of TMP as DHFR inhibitors are highlighted.
Sulfa and trimethoprim-like drugs – antimetabolites acting as carbonic anhydrase, dihydropteroate synthase and dihydrofolate reductase inhibitors
TLDR
The state of the art in the field of antibacterials based on inhibitors of these three enzyme families is revue, when several X-ray crystal structures of such enzymes in complex with their inhibitors were reported.
Charged Propargyl-Linked Antifolates Reveal Mechanisms of Antifolate Resistance and Inhibit Trimethoprim-Resistant MRSA Strains Possessing Clinically Relevant Mutations.
TLDR
A promising lead series is identified that displays significantly greater activity against these mutant enzymes and strains than TMP and employs a series of crystal structures of several mutant enzymes bound to the same inhibitor, revealing a more detailed molecular understanding of drug resistance in this important enzyme.
Antifolates as effective antimicrobial agents: new generations of trimethoprim analogs
TLDR
An historical overview of the attempts to develop drugs that target the folate pathway is presented along with a discussion of the basis of insensitivity to trimethoprim, and the propargyl-linked antifolates are developed as potent inhibitors of TMP-insensitive enzymes and strains.
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A single amino acid substitution in Staphylococcus aureus dihydrofolate reductase determines trimethoprim resistance.
TLDR
Critical evidence concerning the resistance mechanism has been provided by NMR spectral analyses of 15N-labelled TMP in the ternary complexes of both wild-type and mutant enzyme, and studies show that the mutation results in loss of a hydrogen bond between the 4-amino group of TMP and the carbonyl oxygen of Leu5.
Characterization of the gene for the chromosomal dihydrofolate reductase (DHFR) of Staphylococcus epidermidis ATCC 14990: the origin of the trimethoprim-resistant S1 DHFR from Staphylococcus aureus?
TLDR
Site-directed mutagenesis and kinetic analysis of the purified enzymes suggest that a single Phe-->Tyr change at position 98 is the major determinant of trimethoprim resistance.
Structural characteristics of antifolate dihydrofolate reductase enzyme interactions
The ubiquitous enzyme dihydrofolate reductase (DHFR) is responsible for the reduction of 5,6-dihydrofolate to 5,6,7,8-tetrahydrofolate in an NADPH-dependent manner. It is also a key pharmacological
Improving protein solubility through rationally designed amino acid replacements: solubilization of the trimethoprim-resistant type S1 dihydrofolate reductase.
TLDR
A biochemically similar but soluble derivative of the type S1 DHFR that, after production in E.coli, resulted in a 264-fold increase in DHFR activity.
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TLDR
A series of 7,8-dialkylpyrrolo[3,2-f]quinazolines prepared as inhibitors of dihydrofolate reductase (DHFR) were designed to be relatively small and compact, and showed significant distribution into lung and brain tissue.
Characterization of the gene for chromosomal trimethoprim-sensitive dihydrofolate reductase of Staphylococcus aureus ATCC 25923
TLDR
The gene for the trimethoprim-sensitive (Tmps) chromosomal dihydrofolate reductase (DHFR) of Staphylococcus aureus ATCC 25923 was cloned and characterized, with most of the recombinant protein occurring in a soluble and an active form.
Characterization of a staphylococcal trimethoprim resistance gene and its product.
TLDR
The staphylococcal Tp gene codes for a single protein with DHFR activity that appears to be unrelated to DHFR genes that mediate Tp in members of the Enterobacteriaceae.
Identical genes for trimethoprim‐resistant dihydrofolate reductase from Staphylococcus aureus in Australia and Central Europe
TLDR
The nucleotide sequence of a 1.25 kb BglII/EcoRI fragment from the 34 kb trimethoprim (Tp)‐resistant plasmid pABU1 of Staphylococcus aureus 157/4696, isolated in Zürich, was determined and suggests the worldwide dissemination of Tn4OO3 in multiresistant StAPHylococci.
Structural characteristics of small-molecule antifolate compounds
Because of the importance of dihydrofolate reductase (DHFR) inhibitors in the treatment of infections and cancer, a wealth of structural information has been collected. Small-molecule crystallography
Effect of a single amino acid substitution on Escherichia coli dihydrofolate reductase catalysis and ligand binding.
The two isozymes of dihydrofolate reductase (Forms 1 and 2) from, a Trimethoprim-resistant strain of Escherichia coli (RT500) were separated and purified to homogeneity using a simple procedure based
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