| pubmed-article:10886 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:10886 | lifeskim:mentions | umls-concept:C0022940 | lld:lifeskim |
| pubmed-article:10886 | lifeskim:mentions | umls-concept:C0080194 | lld:lifeskim |
| pubmed-article:10886 | lifeskim:mentions | umls-concept:C0039667 | lld:lifeskim |
| pubmed-article:10886 | lifeskim:mentions | umls-concept:C1998793 | lld:lifeskim |
| pubmed-article:10886 | lifeskim:mentions | umls-concept:C1880022 | lld:lifeskim |
| pubmed-article:10886 | pubmed:issue | 3 | lld:pubmed |
| pubmed-article:10886 | pubmed:dateCreated | 1976-12-23 | lld:pubmed |
| pubmed-article:10886 | pubmed:abstractText | Dihydrofolate reductase has been purified from a methotrexate-resistant strain of Lactobacillus casei NCB 6375. By careful attention to growth conditions, up to 2.5 g of enzyme is obtained from a 400 litre culture. The purification procedure, involving poly-ethyleneimine treatment, DEAE-cellulose chromatography and affinity chromatography on methotrexate-aminohexyl-Sepharose, operates on the gram scale, with overall yields of 50-60%. Elution of the affinity column by reverse (upward) flow was used, as it led to recovery of the enzyme in a much smaller volume. The enzyme obtained appears to be more than 98% pure, as judged by gel electrophoresis, isoelectric focusing, and gel filtration. It has a mol.wt. of approx. 17900 and a turnover number of 4s-1 (50mM-triethanolamine/400mM-KCl, pH 7.2, 25 degrees C) with dihydrofolate and NADPH as substrates. The turnover number for folate is 0.02s-1. Michaelis constants for a variety of substrates have been measured by using a new fluorimetric assay (0.36 muM-dihydrofolate; 0.78 muM-NADPH), and binding constants determined by using the quenching of protein fluorescence (dihydrofolate, 2.25 X 10(6)M-1; NADPH, greater than 10(8)M-1). The pH/activity profile shows a single maximum at pH 7.3; at this pH, marked activation by 0.5M-NaCl is observed. | lld:pubmed |
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| pubmed-article:10886 | pubmed:language | eng | lld:pubmed |
| pubmed-article:10886 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:10886 | pubmed:citationSubset | IM | lld:pubmed |
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| pubmed-article:10886 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:10886 | pubmed:month | Sep | lld:pubmed |
| pubmed-article:10886 | pubmed:issn | 0264-6021 | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:RobertsG CGC | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:KingR WRW | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:BurgenA SAS | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:TurnerP CPC | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:DannJ GJG | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:BjurR ARA | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:ScudderPP | lld:pubmed |
| pubmed-article:10886 | pubmed:author | pubmed-author:OstlerGG | lld:pubmed |
| pubmed-article:10886 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:10886 | pubmed:day | 1 | lld:pubmed |
| pubmed-article:10886 | pubmed:volume | 157 | lld:pubmed |
| pubmed-article:10886 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:10886 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:10886 | pubmed:pagination | 559-71 | lld:pubmed |
| pubmed-article:10886 | pubmed:dateRevised | 2009-11-18 | lld:pubmed |
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| pubmed-article:10886 | pubmed:year | 1976 | lld:pubmed |
| pubmed-article:10886 | pubmed:articleTitle | Large-scale purification and characterization of dihydrofolate reductase from a methotrexate-resistant strain of Lactobacillus casei. | lld:pubmed |
| pubmed-article:10886 | pubmed:publicationType | Journal Article | lld:pubmed |
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