Linked Life Data

6608524

Source:http://linkedlifedata.com/resource/pubmed/id/6608524

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
1984-5-4
pubmed:abstractText
An NAD-dependent 5,10-methylenetetrahydrofolate dehydrogenase has been purified to homogeneity from autotrophically and heterotrophically grown cells of Acetobacterium woodii. The enzymes from the differently grown cells were indistinguishable by gel filtration and sodium dodecyl sulfate electrophoresis and have a final specific activity of 670 units mg-1. The enzyme is oxygen-labile; therefore, it was isolated under anaerobic conditions in the presence of dithiothreitol. The oxidized enzyme can be reactivated with 5 mM dithiothreitol, the half-time of activation being 19 min. The forward and reverse reaction initial velocity kinetics was studied and the enzyme was found to follow a substituted (ping-pong) reaction mechanism. With this model, the Km values for NAD and 5,10-methylenetetrahydrofolate are 4.0 and 0.26 mM, while for NADH and 5,10-methenyltetrahydrofolate, they are 2.0 and 1.0 mM, respectively. The equilibrium constant at pH 6.7, determined by the Haldane relationship, is approximately equal to 2.0, favoring the formation of NADH and 5,10-methenyltetrahydrofolate. The purified enzyme is a Mr = 55,000 dimer which lacks 10-formyltetrahydrofolate synthetase and 5,10-methenyltetrahydrofolate cyclohydrolase activities. At pH 6.7, the conversion of 5,10-methylenetetrahydrofolate to 5,10-methenyltetrahydrofolate occurs at a rate of 98,600 mol min-1 mol-1 of enzyme, while the reverse reaction occurs at a rate of 95,600 mol min-1 mol-1 of enzyme.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Mar
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
25
pubmed:volume
259
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
3499-503
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1984
pubmed:articleTitle
Purification and properties of NAD-dependent 5,10-methylenetetrahydrofolate dehydrogenase from Acetobacterium woodii.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't