scispace - formally typeset
Search or ask a question
Topic

Dihydrofolate reductase

About: Dihydrofolate reductase is a research topic. Over the lifetime, 4742 publications have been published within this topic receiving 166010 citations. The topic is also known as: dihydrofolate reductase & uniprot:P00374.


Papers
More filters
Journal ArticleDOI
01 Jan 1988-Proteins
TL;DR: The differences between the active site of all‐atom minimized structure and the experimental structure are similar to differences observed between crystal structures of the same protein.
Abstract: A study of the binding of the antibacterial agent trimethoprim to Escherichia coli dihydrofolate reductase was carried out using energy minimization techniques with both a full, all-atom valence force field and a united atom force field. Convergence criteria ensured that no significant structural or energetic changes would occur with further minimization. Root-mean-square (RMS) deviations of both minimized structures with the experimental structure with the experimental structure were calculated for selected regions of the protein. In the active site, the all-atom minimized structure fit the experimental structure much better than did the united atom structure. To ascertain what constitutes a good fit, the RMS deviations between crystal structures of the same enzyme either from different species or in different crystal environments were compared. The differences between the active site of all-atom minimized structure and the experimental structure are similar to differences observed between crystal structures of the same protein. Finally, the energetics of ligand binding were analyzed for the all-atom minimized coordinates. Strain energy induced in the ligand, the corresponding entropy loss due to shifts in harmonic frequencies, and the role of specific residues in ligand binding were examined. Water molecules, even those not in direct contact with the ligand, were found to have significant interaction energies with the ligand. Thus, the inclusion of at least one shell of waters may be vital for accurate simulations of enzyme complexes.

1,812 citations

Journal ArticleDOI
TL;DR: Mutants of Chinese hamster ovary cells lacking dihydrofolate reductase were isolated after mutagenesis and exposure to high-specific-activity [3H]deoxyuridine as a selective agent.
Abstract: Mutants of Chinese hamster ovary cells lacking dihydrofolate reductase (tetrahydrofolate dehydrogenase, 7,8-dihydrofolate:NADP+ oxidoreductase; EC 1.5.1.3) activity were isolated after mutagenesis and exposure to high-specific-activity [3H]deoxyuridine as a selective agent. Fully deficient mutants could not be isolated starting with wild-type cells, but could readily be selected from a putative heterozygote that contains half of the wild-type level of dihydrofolate reductase activity. The heterozygote itself was selected from wild-type cells by using [3H]deoxyuridine together with methotrexate to reduce intracellular dihydrofolate reductase activity. Fully deficient mutants require glycine, a purine, and thymidine for growth; this phenotype is recessive to wild type in cell hybrids. Revertants have been isolated, one of which produces a heat-labile dihydrofolate reductase activity. These mutants may be useful for metabolic studies relating to cancer chemotherapy and for fine-structure genetic mapping of mutations by using available molecular probes for this gene.

1,421 citations

Journal ArticleDOI
23 Oct 1987-Cell
TL;DR: A dramatic difference in the efficiency of removal of UV-induced pyrimidine dimers from the transcribed and nontranscribed strands of the dihydrofolate reductase (DHFR) gene in cultured hamster and human cells is found and in the 5' flanking region of the human DHFR gene, selective rapid repair occurs in the opposite DNA strand relative to the transcribing strand of the DHFR genes.

1,188 citations

Journal ArticleDOI
TL;DR: A general purification method for recombinant proteins based upon the selective interaction between a poly-histidine peptide, which is fused to the protein of interest, and a novel metal chelate adsorbent is described.
Abstract: We describe a general purification method for recombinant proteins based upon the selective interaction between a poly-histidine peptide, which is fused to the protein of interest, and a novel metal chelate adsorbent. The principle of the technique is illustrated with mouse dihydrofolate reductase. DNA elements coding for adjacent histidines were fused to the mouse dihydrofolate reductase gene. Subsequent expression in E. coli resulted in the production of hybrid proteins that could be purified by immobilized metal ion affinity chromatography, followed by removal of the histidine affinity peptide with carboxypeptidase A.

1,112 citations

Journal ArticleDOI
TL;DR: It is concluded that selective multiplication of the dihydrofolate reductase gene can account for the overproduction of dihydRO- in parental and methotrexate-resistant lines of L1210 murine lymphoma cells.

886 citations


Network Information
Related Topics (5)
Peptide sequence
84.1K papers, 4.3M citations
88% related
Kinase
65.8K papers, 3.5M citations
86% related
DNA
107.1K papers, 4.7M citations
86% related
Phosphorylation
69.3K papers, 3.8M citations
85% related
RNA
111.6K papers, 5.4M citations
85% related
Performance
Metrics
No. of papers in the topic in previous years
YearPapers
202373
2022123
202182
202074
201979
201863