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Documentos Principais

D6566

Sigma-Aldrich

Dihydrofolate Reductase human

≥80% (SDS-PAGE), recombinant, expressed in E. coli, ≥1 units/mg protein

Sinônimo(s):

DHFR, Tetrahydrofolate NADP+ oxidoreductase

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About This Item

Número da licença da enzima:
Número MDL:
Código UNSPSC:
12352204
NACRES:
NA.54

recombinante

expressed in E. coli

Nível de qualidade

Ensaio

≥80% (SDS-PAGE)

Formulário

solution

atividade específica

≥1 units/mg protein

peso molecular

25 kDa

concentração

0.02-0.06 mg/mL

nº de adesão UniProt

Condições de expedição

wet ice

temperatura de armazenamento

−20°C

Informações sobre genes

human ... DHFR(1719)

Descrição geral

Human DHFR is an 186 amino acid protein with an apparent molecular weight of 25 kDa. It is 30% homologous to the E. coli protein and up to 70% homologous to vertebrate proteins.

Aplicação

Dihydrofolate Reductase human has been used:
  • to investigate the stable expression of green fluorescent protein and the targeted disruption of thioredoxin peroxidase-1 gene in Babesia bovis
  • to study the structural analysis of human dihydrofolate reductase as a binary complex
  • to study its in vitro kinetic assay for the enzyme inhibition study
Human dihydrofolate reductase has been used in a study to investigate the stable expression of green fluorescent protein and the targeted disruption of thioredoxin peroxidase-1 gene in Babesia bovis. Human dihydrofolate reductase has also been used in a study to investigate the structural analysis of human dihydrofolate reductase as a binary complex.

Ações bioquímicas/fisiológicas

Dihydrofolate reductase (DHFR) is a key enzyme in thymidine synthesis. It catalyzes the reduction of dihydrofolate (DHF) to tetrahydrofolate (THF). At a much lower rate, it catayzes the conversion of folate to THF. Since thymidine is a necessary substrate for DNA synthesis, DHFR is a target for anticancer drug development. Methotrexate is the prototype dihydrofolate reductase inhibitor. The enzyme from Sigma has been used in the inhibitory studies of Leishmaniasis donovani pteridine reductase 1 (PTR1). The enzyme has also been used as a positive control to measure the DHFR activity of a protein, MS0308, purified from Mycobacterium smegmatis.
Km5,6
NADPH 0.16 mM
7,8-dihydrofolate 0.03 mM
8-methylpterin 0.13 mM
Ki7
Folate 2.6x10-5 mM
Methotrexate 6.1-9x10-9
The human DHFR gene, as well as DHFR genes in other mammalian species, overcome the inhibitory effects of methotrexate by a mechanism of gene amplification or by amino-acid mutagenesis. Dihydroflate reductase (DHFR) catalyzes the NADPH dependent reduction of dihydrofolate (DHF) to tetrahydrofolate (THF) and, at a much lower rate, the conversion of folate to THF. The reaction product, THF, is an essential cofactor in the conversion of deoxyuridylate (dUMP) to deoxythymidylate (dTMP) by thymidylate synthetase. It is a key enzyme in thymidine synthesis. Therefore, DHFR is a critical enzyme in DNA synthesis and has become a target for drug development and cancer therapy. The variations between DHFR from different sources have enabled the development of species selective DHFR inhibitors, such as trimethoprim (antibacterial and antifungal), pyrimethamine (antiprotozoal), and methotrexate; MTX (antineoplastic, antipsoriatic, and anti-inflammatory).

Definição da unidade

One unit will convert 1.0 μmole of dihydrofolic acid to tetrahydrofolic acid in 1 minute at pH 7.5 at 22 °C.

forma física

Solution in 10 mM Tris pH 8, 1 mM EDTA, 0.5 mM DTT, 5 μM NADPH, protease inhibitors, and 50% glycerol.

substrato

Código de classe de armazenamento

12 - Non Combustible Liquids

Classe de risco de água (WGK)

WGK 1

Ponto de fulgor (°F)

Not applicable

Ponto de fulgor (°C)

Not applicable

Equipamento de proteção individual

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


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Masahito Asada et al.
Molecular and biochemical parasitology, 181(2), 162-170 (2011-11-24)
We have achieved stable expression of green fluorescent protein (GFP) in Babesia bovis by using the WR99210/human dihydrofolate reductase (DHFR) gene selection system. A GFP-expression plasmid with a dhfr expression cassette (DHFR-gfp) was constructed and transfected into B. bovis by
Eukaryotic dihydrofolate reductase.
R L Blakley
Advances in enzymology and related areas of molecular biology, 70, 23-102 (1995-01-01)
Dimitrios Evangelopoulos et al.
The FEBS journal, 278(24), 4824-4832 (2011-10-07)
Mycobacterium tuberculosis, the most successful bacterial pathogen, causes tuberculosis, a disease that still causes more than 2 million deaths per year. Arylamine N-acetyltransferase is an enzyme that is conserved in most Mycobacterium spp. The nat gene belongs to an operon that
Eyal Sharon et al.
Proceedings of the National Academy of Sciences of the United States of America, 111(1), 451-456 (2013-12-18)
E2F transcription factors play pivotal roles in controlling the expression of genes involved in cell-cycle progression. Different viruses affect E2F1/retinoblastoma (Rb) interactions by diverse mechanisms releasing E2F1 from its suppressor Rb, enabling viral replication. We show that in T cells
Samuel Genheden
Journal of chemical information and modeling, 52(11), 3013-3021 (2012-11-02)
In this paper, I evaluate the usefulness of protein homology models in rigorous free-energy simulations to determine ligand affinities. Two templates were used to create models of the factor Xa protein and one template was used for dihydrofolate reductase from

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