Linked Life Data

10811639

Source:http://linkedlifedata.com/resource/pubmed/id/10811639

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
32
pubmed:dateCreated
2000-9-14
pubmed:databankReference
pubmed:abstractText
Chain elongation of fatty acids is an important cellular process and is believed to occur in the endoplasmic reticulum of all eukaroytic cells. Herein we describe the cloning and characterization of a peroxisomal NADPH-specific trans-2-enoyl-CoA reductase, the key enzyme for a proposed peroxisomal chain elongation pathway. The reductase was solubilized and partially purified from guinea pig liver peroxisomes by affinity chromatography. On SDS-polyacrylamide gel electrophoresis, a 40-kDa band was identified as the enzyme, and its partial amino acid sequence (27 amino acids) was determined. A full-length cDNA for the reductase was cloned from a guinea pig liver cDNA library. The open reading frame of this nucleotide sequence encodes a 302-amino acid polypeptide with a calculated molecular mass of 32.5 kDa. Full-length mouse and human cDNA clones encoding homologous proteins have also been isolated. All of these translated polypeptides have the type I peroxisomal targeting signal, AKL, at the carboxyl terminus. The identity of the cloned enoyl-CoA reductase cDNAs was confirmed by expressing the guinea pig and human cDNAs in Escherichia coli. The His-tagged recombinant enzymes were found to have very high NADPH-specific 2-enoyl-CoA reductase activity with similar properties and specificity as the liver peroxisomal reductase. Both the natural and the recombinant enzyme catalyze the reduction of trans-2-enoyl-CoAs of varying chain lengths from 6:1 to 16:1, having maximum activity with 10:1 CoA. Northern blot analysis demonstrated that a single transcript of 1.3 kilobases is present in most mouse tissues, with particularly high concentrations in liver and kidney.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
11
pubmed:volume
275
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
24333-40
pubmed:dateRevised
2009-11-19
pubmed:meshHeading
pubmed-meshheading:10811639-Amino Acid Sequence, pubmed-meshheading:10811639-Animals, pubmed-meshheading:10811639-Base Sequence, pubmed-meshheading:10811639-Chromatography, Affinity, pubmed-meshheading:10811639-Cloning, Molecular, pubmed-meshheading:10811639-Fatty Acid Desaturases, pubmed-meshheading:10811639-Fatty Acid Synthetase Complex, pubmed-meshheading:10811639-Guinea Pigs, pubmed-meshheading:10811639-Humans, pubmed-meshheading:10811639-Kinetics, pubmed-meshheading:10811639-Liver, pubmed-meshheading:10811639-Mammals, pubmed-meshheading:10811639-Mice, pubmed-meshheading:10811639-Microsomes, Liver, pubmed-meshheading:10811639-Molecular Sequence Data, pubmed-meshheading:10811639-NADH, NADPH Oxidoreductases, pubmed-meshheading:10811639-Peroxisomes, pubmed-meshheading:10811639-Rats, pubmed-meshheading:10811639-Recombinant Proteins, pubmed-meshheading:10811639-Sequence Alignment, pubmed-meshheading:10811639-Sequence Homology, Amino Acid, pubmed-meshheading:10811639-Substrate Specificity
pubmed:year
2000
pubmed:articleTitle
Molecular cloning and expression of mammalian peroxisomal trans-2-enoyl-coenzyme A reductase cDNAs.
pubmed:affiliation
Mental Health Research Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48104-1687, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.