Interaction of methotrexate, folates, and pyridine nucleotides with dihydrofolate reductase: calorimetric and spectroscopic binding studies.

Subramanian, S.; Kaufman, Bernard T.

doi:10.1073/pnas.75.7.3201

Cited by 43 publications

(25 citation statements)

References 25 publications

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“…Being structural analogs of the substrate folate, these drugs competitively inhibit the enzyme. In methotrexate, an amino group replaces the 4-hydroxyl group of folate [ 16 , 19 - 23 ]. The active site of DHFR comprises the amino acid residues Ile-7, Leu-22, Phe-31, Phe-34, Arg-70, Val-115 and Tyr121.…”

Section: Introductionmentioning

confidence: 99%

“…Of the eight strands seven are parallel and the eighth runs antiparallel. Successive β strands are connected by four α helices [ 19 , 22 ]. A major sub domain surrounding the active site contains a loop of residues 9- 24 called “loop 1” (also termed “Met20 loop”).…”

Section: Introductionmentioning

confidence: 99%

“…The Met20 and other loops near the active site are highly flexible and promoting the release of the tetrahydrofolate product after the catalytic reduction of dihydrofolate by NADPH [ 16 ]. The nicotinamide ring of the NADPH is stabilized by the Met20 loop which promotes the transfer of hydride from NADPH to dihydrofolate [ 19 , 24 ].…”

Section: Introductionmentioning

confidence: 99%

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A comparative molecular docking study of curcumin and methotrexate to dihydrofolate reductase

2017

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Interaction of curcumin (CUR) with the enzyme dihydrofolate reductase (DHFR) was studied by molecular docking using AutoDock 4.2 as the docking software application. AutoDock 4.2 software serves as a valid and acceptable docking application to study the interactions of small compounds with proteins. Interactions of curcumin with DHFR were compared to those of methotrexate (MTX), a known inhibitor of the enzyme. The calculated free energy of binding (ΔG binding) shows that curcumin (ΔG = -9.02 kcal/mol; Ki = 243 nM) binds with affinity comparable to or better than MTX (ΔG = -8.78 kcal/mol; Ki = 363 nM). Binding interactions of curcumin with active site residues of the enzyme are also predicted. Curcumin appears to bind in a bent conformation making extensive VDW contacts in the active site of the enzyme. Hydrogen bonding and pi-pi interaction with key active site residues are also observed. Thus, curcumin can be considered as a good lead compound in the development of new inhibitors of DHFR, which is a potential target of anti-cancer drugs. The results of these studies can serve as a starting point for further computational and experimental studies.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A comparative molecular docking study of curcumin and methotrexate to dihydrofolate reductase

2017

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“…The dissociation constant of MTX from its ternary complex with DHFR is on the order of 10 10 (Hood and Roberts, 1978), which corresponds to about 14 kcallmo1e of binding energy (see also Subramanian and Kaufman, 1978). Based on structural information, that overall energy of binding must arise from a composite of many different kinds of intermolecular interaction.…”

Section: Dhfr-methotrexate Complexesmentioning

confidence: 99%

Protein Design and the Development of New Therapeutics and Vaccines

Hook¹,

Poste²,

Smith³

1990

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“…Co-elution of MTX with Its Protein Target DHFR-As a first proof of principle, we monitored the co-elution of the folate antagonist MTX with its high affinity (K d ϭ 4.8 nM) enzyme target, DHFR (24). The MTX-DHFR complex represents a well characterized drug-target interaction supported by extensive biophysical and biological data addressing fundamental structure activity relationships (25,26).…”

Section: Concept: Target Identification By Chromatographic Co-elution-mentioning

confidence: 99%

Target Identification by Chromatographic Co-elution: Monitoring of Drug-Protein Interactions without Immobilization or Chemical Derivatization

Chan

Vuckovic²,

Sleno³

et al. 2012

Molecular & Cellular Proteomics

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Bioactive molecules typically mediate their biological effects through direct physical association with one or more cellular proteins. The detection of drug-target interactions is therefore essential for the characterization of compound mechanism of action and off-target effects, but generic label-free approaches for detecting binding events in biological mixtures have remained elusive. Here, we report a method termed target identification by chromatographic co-elution (TICC) for routinely monitoring the interaction of drugs with cellular proteins under nearly physiological conditions in vitro based on simple liquid chromatographic separations of cell-free lysates. Correlative proteomic analysis of drug-bound protein fractions by shotgun sequencing is then performed to identify candidate target(s). The method is highly reproducible, does not require immobilization or derivatization of drug or protein, and is applicable to diverse natural products and synthetic compounds. The capability of TICC to detect known drug-protein target physical interactions (K d range: micromolar to nanomolar) is demonstrated both qualitatively and quantitatively. We subsequently used TICC to uncover the sterol biosynthetic enzyme Erg6p as a novel putative anti-fungal target. Furthermore, TICC identified Asc1 and Dak1, a core 40 S ribosomal protein that represses gene expression, and dihydroxyacetone kinase involved in stress adaptation, respectively, as novel yeast targets of a dopamine receptor agonist.

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Interaction of methotrexate, folates, and pyridine nucleotides with dihydrofolate reductase: calorimetric and spectroscopic binding studies.

Abstract: The thermodynamic parameters, AG, AH, and AS, characterizing the tight binding of methotrexate, folates, and pyridine nucleotides to chicken liver dihydrofolate reductase (5,6,7,8-

Cited by 43 publications

References 25 publications

A comparative molecular docking study of curcumin and methotrexate to dihydrofolate reductase

A comparative molecular docking study of curcumin and methotrexate to dihydrofolate reductase

Protein Design and the Development of New Therapeutics and Vaccines

Target Identification by Chromatographic Co-elution: Monitoring of Drug-Protein Interactions without Immobilization or Chemical Derivatization

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