scholarly journals Chimeric dihydrofolate reductases display properties of modularity and biophysical diversity

2019 ◽  
Author(s):  
João V. Rodrigues ◽  
C. Brandon Ogbunugafor ◽  
Daniel L. Hartl ◽  
Eugene I. Shakhnovich
Keyword(s):  
Dihydrofolate Reductases

scholarly journals Yeast-based automated high-throughput screens to identify anti-parasitic lead compounds

Open Biology ◽  
2013 ◽  
Vol 3 (2) ◽  
pp. 120158 ◽  
Author(s):  
Elizabeth Bilsland ◽  
Andrew Sparkes ◽  
Kevin Williams ◽  
Harry J. Moss ◽  
Michaela de Clare ◽  
...  
Keyword(s):  
Side Effects ◽  
Trypanosoma Brucei ◽  
Fluorescent Proteins ◽  
Screening Method ◽  
Lead Compounds ◽  
Dihydrofolate Reductases ◽  
Parasitic Activity ◽  
High Throughput Screens ◽  
Specific Inhibitors

We have developed a robust, fully automated anti-parasitic drug-screening method that selects compounds specifically targeting parasite enzymes and not their host counterparts, thus allowing the early elimination of compounds with potential side effects. Our yeast system permits multiple parasite targets to be assayed in parallel owing to the strains’ expression of different fluorescent proteins. A strain expressing the human target is included in the multiplexed screen to exclude compounds that do not discriminate between host and parasite enzymes. This form of assay has the advantages of using known targets and not requiring the in vitro culture of parasites. We performed automated screens for inhibitors of parasite dihydrofolate reductases, N -myristoyltransferases and phosphoglycerate kinases, finding specific inhibitors of parasite targets. We found that our ‘hits’ have significant structural similarities to compounds with in vitro anti-parasitic activity, validating our screens and suggesting targets for hits identified in parasite-based assays. Finally, we demonstrate a 60 per cent success rate for our hit compounds in killing or severely inhibiting the growth of Trypanosoma brucei , the causative agent of African sleeping sickness.

Dihydrofolate reductases of Escherichia coli and bacteriophage T4. Spectrofluorometric study

Biochemistry ◽  
1973 ◽  
Vol 12 (3) ◽  
pp. 372-380 ◽  
Author(s):  
John S. Erickson ◽  
Christopher K. Mathews
Keyword(s):  
Escherichia Coli ◽  
Bacteriophage T4 ◽  
Dihydrofolate Reductases

Investigation of Osmolyte Effects on FolM: Comparison with Other Dihydrofolate Reductases

Biochemistry ◽  
2014 ◽  
Vol 53 (8) ◽  
pp. 1330-1341 ◽  
Author(s):  
Purva P. Bhojane ◽  
Michael R. Duff ◽  
Harini C. Patel ◽  
Melissa E. Vogt ◽  
Elizabeth E. Howell
Keyword(s):  
Dihydrofolate Reductases

scholarly journals Dual-Target Inhibitors of the Folate Pathway Inhibit Intrinsically Trimethoprim-Resistant DfrB Dihydrofolate Reductases

2020 ◽  
Vol 11 (11) ◽  
pp. 2261-2267
Author(s):  
Jacynthe L. Toulouse ◽  
Genbin Shi ◽  
Claudèle Lemay-St-Denis ◽  
Maximilian C. C. J. C. Ebert ◽  
Daniel Deon ◽  
...  
Keyword(s):  
Folate Pathway ◽  
Dihydrofolate Reductases ◽  
Dual Target

scholarly journals Crystal Structures of Klebsiella pneumoniae Dihydrofolate Reductase Bound to Propargyl-Linked Antifolates Reveal Features for Potency and Selectivity

2014 ◽  
Vol 58 (12) ◽  
pp. 7484-7491 ◽  
Author(s):  
Kristen M. Lamb ◽  
Michael N. Lombardo ◽  
Jeremy Alverson ◽  
Nigel D. Priestley ◽  
Dennis L. Wright ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Crystal Structures ◽  
Dihydrofolate Reductase ◽  
Next Generation ◽  
Gram Negative ◽  
Content Type ◽  
Resistant Species ◽  
Loop Conformations ◽  
The Family ◽  
Dihydrofolate Reductases

ABSTRACTResistance to the antibacterial antifolate trimethoprim (TMP) is increasing in members of the familyEnterobacteriaceae, driving the design of next-generation antifolates effective against these Gram-negative pathogens. The propargyl-linked antifolates are potent inhibitors of dihydrofolate reductases (DHFR) from several TMP-sensitive and -resistant species, includingKlebsiella pneumoniae. Recently, we have determined that these antifolates inhibit the growth of strains ofK. pneumoniae, some with MIC values of 1 μg/ml. In order to further the design of potent and selective antifolates against members of theEnterobacteriaceae, we determined the first crystal structures ofK. pneumoniaeDHFR bound to two of the propargyl-linked antifolates. These structures highlight that interactions with Leu 28, Ile 50, Ile 94, and Leu 54 are necessary for potency; comparison with structures of human DHFR bound to the same inhibitors reveal differences in residues (N64E, P61G, F31L, and V115I) and loop conformations (residues 49 to 53) that may be exploited for selectivity.

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