Document Detail


Dihydrofolate reductase from bovine liver. Enzymatic and structural properties.
MedLine Citation:
PMID:  1204646     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Dihydrofolate reductase from bovine liver has been purified 5000-fold employing conventional techniques and methotrexate/aminohexyl/Sepharose affinity chromatography. Electrophoresis of the isolated enzyme on polyacrylamide gels resulted in the separation of two enzymatically active protein components which were not interconvertible by treatment with dihydrofolate and/or the coenzyme. The two forms, present in a ratio of 20:1, were found by isoelectric focusing to have isoelectric points of 7.15 and 5.94. They had identical specific activities toward dihydrofolate (26.1-27.0 U/mg) and folate (1.3-2.2 U/mg), and had identical molecular weights (23500) and amino acid compositions. Due to the small quantity of the acidic form and the similarity of the two forms, the amino-terminal sequence (19 residues) was determined on a mixture of carboxymethylated reductase. The single sulfhydryl group of the enzyme can be modified by several sulfhydryl reagents in the native enzyme without loss of activity. Modification of the same residue occurs in the denaturated state and partially inhibits renaturation to the fully acitve enzyme. One disulfide bridge was detected by reduction and alkylation. The cleavage of this bond did not effect the enzymatic activity.
Authors:
H Baumann; K J Wilson
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  European journal of biochemistry / FEBS     Volume:  60     ISSN:  0014-2956     ISO Abbreviation:  Eur. J. Biochem.     Publication Date:  1975 Dec 
Date Detail:
Created Date:  1976-03-18     Completed Date:  1976-03-18     Revised Date:  2009-10-27    
Medline Journal Info:
Nlm Unique ID:  0107600     Medline TA:  Eur J Biochem     Country:  GERMANY, WEST    
Other Details:
Languages:  eng     Pagination:  9-15     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Amino Acids / analysis
Animals
Cattle
Chromatography, Affinity
Isoenzymes / isolation & purification,  metabolism
Liver / enzymology*
Molecular Weight
Protein Binding
Species Specificity
Streptococcus / enzymology
Sulfhydryl Reagents / pharmacology
Temperature
Tetrahydrofolate Dehydrogenase / isolation & purification,  metabolism*
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Isoenzymes; 0/Sulfhydryl Reagents; EC 1.5.1.3/Tetrahydrofolate Dehydrogenase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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