Linked Life Data

1958662

Source:http://linkedlifedata.com/resource/pubmed/id/1958662

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
47
pubmed:dateCreated
1992-1-3
pubmed:abstractText
The LAT1 gene encoding the dihydrolipoamide acetyltransferase component (E2) of the pyruvate dehydrogenase (PDH) complex from Saccharomyces cerevisiae was disrupted, and the lat1 null mutant was used to analyze the structure and function of the domains of E2. Disruption of LAT1 did not affect the viability of the cells. Apparently, flux through the PDH complex is not required for growth of S. cerevisiae under the conditions tested. The wild-type and mutant PDH complexes were purified to near-homogeneity and were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and enzyme assays. Mutant cells transformed with LAT1 on a unit-copy plasmid produced a PDH complex very similar to that of the wild-type PDH complex. Deletion of most of the putative lipoyl domain (residues 8-84) resulted in loss of about 85% of the overall activity, but did not affect the acetyltransferase activity of E2 or the binding of pyruvate dehydrogenase (E1), dihydrolipoamide dehydrogenase (E3), and protein X to the truncated E2. Similar results were obtained by deleting the lipoyl domain plus the first hinge region (residues 8-145) and by replacing lysine-47, the putative site of covalent attachment of the lipoyl moiety, by arginine. Although the lipoyl domain of E2 and/or its covalently bound lipoyl moiety were removed, the mutant complexes retained 12-15% of the overall activity of the wild-type PDH complex. Replacement of both lysine-47 in E2 and the equivalent lysine-43 in protein X by arginine resulted in complete loss of overall activity of the mutant PDH complex.(ABSTRACT TRUNCATED AT 250 WORDS)
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
26
pubmed:volume
30
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
11249-54
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:1958662-Acetyltransferases, pubmed-meshheading:1958662-Amino Acid Sequence, pubmed-meshheading:1958662-Base Sequence, pubmed-meshheading:1958662-Blotting, Southern, pubmed-meshheading:1958662-Chromosome Deletion, pubmed-meshheading:1958662-Cloning, Molecular, pubmed-meshheading:1958662-DNA, Fungal, pubmed-meshheading:1958662-Dihydrolipoyllysine-Residue Acetyltransferase, pubmed-meshheading:1958662-Escherichia coli, pubmed-meshheading:1958662-Immunoblotting, pubmed-meshheading:1958662-Molecular Sequence Data, pubmed-meshheading:1958662-Oligodeoxyribonucleotides, pubmed-meshheading:1958662-Open Reading Frames, pubmed-meshheading:1958662-Polymerase Chain Reaction, pubmed-meshheading:1958662-Pyruvate Dehydrogenase Complex, pubmed-meshheading:1958662-Recombinant Proteins, pubmed-meshheading:1958662-Restriction Mapping, pubmed-meshheading:1958662-Saccharomyces cerevisiae, pubmed-meshheading:1958662-Saccharomyces cerevisiae Proteins
pubmed:year
1991
pubmed:articleTitle
Functional analysis of the domains of dihydrolipoamide acetyltransferase from Saccharomyces cerevisiae.
pubmed:affiliation
Clayton Foundation Biochemical Institute, University of Texas, Austin 78712.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't