Linked Life Data

1748668

Source:http://linkedlifedata.com/resource/pubmed/id/1748668

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
35
pubmed:dateCreated
1992-1-17
pubmed:databankReference
pubmed:abstractText
We have purified the enzyme 5,10-methylenetetrahydrofolate dehydrogenase (EC 1.5.1.5) from Escherichia coli to homogeneity by a newly devised procedure. The enzyme has been purified at least 2,000-fold in a 31% yield. The specific activity of the enzyme obtained is 7.4 times greater than any previous preparation from this source. The purified enzyme is specific for NADP. The protein also contains 5,10-methenyltetrahydrofolate cyclohydrolase (EC 3.5.4.9) activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and behavior on a molecular sieving column suggest that the enzyme is a dimer of identical subunits. We have cloned the E. coli gene coding for the enzyme through the use of polymerase chain reaction based on primers designed from the NH2 terminal analysis of the isolated enzyme. We sequenced the gene. The derived amino acid sequence of the enzyme contains 287 amino acids of Mr 31,000. The sequence shows 50% identity to two bifunctional mitochondrial enzymes specific for NAD, and 40-45% identity to the presumed dehydrogenase/cyclohydrolase domains of the trifunctional C1-tetrahydrofolate synthase of yeast mitochondria and cytoplasm and human and rat cytoplasm. An identical sequence of 14 amino acids with no gaps is present in all 7 sequences.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
266
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
23953-8
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:1748668-Amino Acid Sequence, pubmed-meshheading:1748668-Aminohydrolases, pubmed-meshheading:1748668-Animals, pubmed-meshheading:1748668-Base Sequence, pubmed-meshheading:1748668-Chromatography, Affinity, pubmed-meshheading:1748668-Chromatography, Ion Exchange, pubmed-meshheading:1748668-Cloning, Molecular, pubmed-meshheading:1748668-Escherichia coli, pubmed-meshheading:1748668-Humans, pubmed-meshheading:1748668-Kinetics, pubmed-meshheading:1748668-Methylenetetrahydrofolate Dehydrogenase (NADP), pubmed-meshheading:1748668-Molecular Sequence Data, pubmed-meshheading:1748668-Multienzyme Complexes, pubmed-meshheading:1748668-NAD, pubmed-meshheading:1748668-Polymerase Chain Reaction, pubmed-meshheading:1748668-Recombinant Proteins, pubmed-meshheading:1748668-Restriction Mapping, pubmed-meshheading:1748668-Sequence Homology, Nucleic Acid, pubmed-meshheading:1748668-Substrate Specificity
pubmed:year
1991
pubmed:articleTitle
Purification, characterization, cloning, and amino acid sequence of the bifunctional enzyme 5,10-methylenetetrahydrofolate dehydrogenase/5,10-methenyltetrahydrofolate cyclohydrolase from Escherichia coli.
pubmed:affiliation
Department of Molecular and Cell Biology, University of California, Berkeley 94720.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, U.S. Gov't, P.H.S.