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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
33
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pubmed:dateCreated |
1991-12-26
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pubmed:databankReference |
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55194,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55195,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55196,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55197,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55198,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55199,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55200,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M55201,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/M74131,
http://linkedlifedata.com/resource/pubmed/xref/GENBANK/S66785
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pubmed:abstractText |
NAD(+)-dependent isocitrate dehydrogenase from Saccharomyces cerevisiae is composed of two nonidentical subunits, designated IDH1 (Mr approximately 40,000) and IDH2 (Mr approximately 39,000). We have isolated and characterized a yeast genomic clone containing the IDH2 gene. The amino acid sequence deduced from the gene indicates that IDH2 is synthesized as a precursor of 369 amino acids (Mr 39,694) and is processed upon mitochondrial import to yield a mature protein of 354 amino acids (Mr 37,755). Amino acid sequence comparison between S. cerevisiae IDH2 and S. cerevisiae NADP(+)-dependent isocitrate dehydrogenase shows no significant sequence identity, whereas comparison of IDH2 and Escherichia coli NADP(+)-dependent isocitrate dehydrogenase reveals a 33% sequence identity. To confirm the identity of the IDH2 gene and examine the relationship between IDH1 and IDH2, the IDH2 gene was disrupted by genomic replacement in a haploid yeast strain. The disruption strain expressed no detectable IDH2, as determined by Western blot analysis, and was found to lack NAD(+)-dependent isocitrate dehydrogenase activity, indicating that IDH2 is essential for a functional enzyme. Overexpression of IDH2, however, did not result in increased NAD(+)-dependent isocitrate dehydrogenase activity, suggesting that both IDH1 and IDH2 subunits are required for catalytic activity. The disruption strain was unable to utilize acetate as a carbon source and exhibited a 2-fold slower growth rate than wild type strains on glycerol or lactate. This growth phenotype is consistent with NAD(+)-dependent isocitrate dehydrogenase performing an essential role in the oxidative function of the citric acid cycle.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Fungal,
http://linkedlifedata.com/resource/pubmed/chemical/Isocitrate Dehydrogenase,
http://linkedlifedata.com/resource/pubmed/chemical/Isoenzymes,
http://linkedlifedata.com/resource/pubmed/chemical/NAD,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins
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pubmed:status |
MEDLINE
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pubmed:month |
Nov
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pubmed:issn |
0021-9258
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
25
|
pubmed:volume |
266
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pubmed:geneSymbol |
IDH2
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pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
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pubmed:pagination |
22199-205
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pubmed:dateRevised |
2007-11-14
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pubmed:meshHeading |
pubmed-meshheading:1939242-Amino Acid Sequence,
pubmed-meshheading:1939242-Base Sequence,
pubmed-meshheading:1939242-Blotting, Southern,
pubmed-meshheading:1939242-Cloning, Molecular,
pubmed-meshheading:1939242-DNA, Fungal,
pubmed-meshheading:1939242-Escherichia coli,
pubmed-meshheading:1939242-Genes, Fungal,
pubmed-meshheading:1939242-Isocitrate Dehydrogenase,
pubmed-meshheading:1939242-Isoenzymes,
pubmed-meshheading:1939242-Mitochondria,
pubmed-meshheading:1939242-Molecular Sequence Data,
pubmed-meshheading:1939242-Molecular Weight,
pubmed-meshheading:1939242-NAD,
pubmed-meshheading:1939242-Recombinant Proteins,
pubmed-meshheading:1939242-Restriction Mapping,
pubmed-meshheading:1939242-Saccharomyces cerevisiae,
pubmed-meshheading:1939242-Sequence Homology, Nucleic Acid
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pubmed:year |
1991
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pubmed:articleTitle |
NAD(+)-dependent isocitrate dehydrogenase. Cloning, nucleotide sequence, and disruption of the IDH2 gene from Saccharomyces cerevisiae.
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pubmed:affiliation |
Department of Biological Chemistry, University of California, Irvine 92717.
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pubmed:publicationType |
Journal Article,
Comparative Study,
Research Support, U.S. Gov't, P.H.S.,
Research Support, Non-U.S. Gov't
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