Subunit structure, expression, and function of NAD(H)-specific isocitrate dehydrogenase in Saccharomyces cerevisiae

Keys, Daniel A.; McAlister-Henn, Lee

doi:10.1128/jb.172.8.4280-4287.1990

Cited by 105 publications

(98 citation statements)

References 40 publications

(29 reference statements)

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“…The molecular properties of NAD-IDH from C. reinhardtii (molecular mass, number of subunits, and configuration of the active site) are similar to those reported from nonphotosynthetic eukaryotes such as S. cerevisiae (Keys and McAlister-Henn, 1990) or pig heart (Giorgio et al, 1970). However, the NAD-IDH from C. reinhardtii did not exhibit the same regulatory properties, since citrate, ADP, and AMP did not act as effectors.…”

Section: Discussionsupporting

confidence: 52%

“…Comparable specific activities have been reported for the purified preparations of NAD-IDH from Phycomyces blakesleeanus (AlvarezVillafañ e et al, 1996) and Saccharomyces cerevisiae (Keys and McAlister-Henn, 1990). …”

Section: Purification Of Nad-idh From C Reinhardtiimentioning

confidence: 73%

“…This enzyme has been purified from several nonphotosynthetic eukaryotes such as fungi (Keys and McAlister-Henn, 1990;AlvarezVillafañ e et al, 1996) and animals (Giorgio et al, 1970), in which it appears to be a 300-kD octamer. Its key regulatory role in the TCA cycle is well documented.…”

mentioning

confidence: 99%

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Purification and Characterization of NAD-Isocitrate Dehydrogenase from Chlamydomonas reinhardtii1

Martínez-Rivas¹,

Vega

1998

Plant Physiology

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NAD-isocitrate dehydrogenase (NAD-IDH) from the eukaryotic microalga Chlamydomonas reinhardtii was purified to electrophoretic homogeneity by successive chromatography steps on Phenyl-Sepharose, Blue-Sepharose, diethylaminoethyl-Sephacel, and Sephacryl S-300 (all Pharmacia Biotech). The 320-kD enzyme was found to be an octamer composed of 45-kD subunits. The presence of isocitrate plus Mn 2؉ protected the enzyme against thermal inactivation or inhibition by specific reagents for arginine or lysine. NADH was a competitive inhibitor (K i , 0.14 mM) and NADPH was a noncompetitive inhibitor (K i , 0.42 mM) with respect to NAD ؉ . Citrate and adenine nucleotides at concentrations less than 1 mM had no effect on the activity, but 10 mM citrate, ATP, or ADP had an inhibitory effect. In addition, NAD-IDH was inhibited by inorganic monovalent anions, but L-amino acids and intermediates of glycolysis and the tricarboxylic acid cycle had no significant effect. These data support the idea that NAD-IDH from photosynthetic organisms may be a key regulatory enzyme within the tricarboxylic acid cycle.

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Section: Discussionsupporting

confidence: 52%

Section: Purification Of Nad-idh From C Reinhardtiimentioning

confidence: 73%

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Purification and Characterization of NAD-Isocitrate Dehydrogenase from Chlamydomonas reinhardtii1

Martínez-Rivas¹,

Vega

1998

Plant Physiology

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“…Immunoblot analyses for yeast proteins were conducted using antisera specific for IDH (1:2000 dilution) [3]; for subunits of cytochrome c oxidase including COX2 (1:2000 dilution), COX3 (1:1000 dilution), and COX4 (1:1000 dilution) (Molecular Probes, Inc.); for β-actin (1:2000 dilution, Abcam), and for CIT2 (1:2000 dilution, kindly provided by Drs. Mark T. McCammon and Gyula Kispal).…”

Section: Western Blot Analysesmentioning

confidence: 99%

“…The enzyme is subject to extensive allosteric control, and the reaction is essentially irreversible under physiological conditions [1,2]. Yeast IDH is an octamer composed of four catalytic IDH2 subunits and four homologous regulatory IDH1 subunits [3,4].…”

Section: Introductionmentioning

confidence: 99%

Suppression of metabolic defects of yeast isocitrate dehydrogenase and aconitase mutants by loss of citrate synthase

Hakala

Weintraub

et al. 2008

Archives of Biochemistry and Biophysics

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Yeast mutants lacking mitochondrial NAD + -specific isocitrate dehydrogenase (idhΔ) or aconitase (aco1 Δ) were found to share several growth phenotypes as well as patterns of specific protein expression that differed from the parental strain. These shared properties of idhΔ and aco1 Δ strains were eliminated or moderated by co-disruption of the CIT1 gene encoding mitochondrial citrate synthase. Gas chromatography/mass spectrometry analyses indicated a particularly dramatic increase in cellular citrate levels in idhΔ and aco1 Δ strains, whereas citrate levels were substantially lower in idhΔcit1 Δ and aco1 Δcit1 Δ strains. Exogenous addition of citrate to parental strain cultures partially recapitulated effects of high endogenous levels of citrate in idh Δ and aco1 Δ strains. Finally, effects of elevated cellular citrate in idhΔ and aco1 Δ mutant strains were partially alleviated by addition of iron or by an increase in pH of the growth medium, suggesting that detrimental effects of citrate are due to elevated levels of the ionized form of this metabolite.

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Isolation of aHistoplasma capsulatum cDNA that complements a mitochondrial NAD(+)-isocitrate dehydrogenase subunit I-deficient mutant ofSaccharomyces cerevisiae

Johnson

McEwen

1999

Yeast

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A cDNA library was prepared from Histoplasma capsulatum strain G‐217B yeast cells and an apparently full‐length cDNA for a subunit of the citric acid cycle enzyme NAD(+)‐isocitrate dehydrogenase was identified by sequence analysis. Its predicted amino acid sequence is more similar to the IDH1 regulatory subunit of S. cerevisiae NAD(+)‐isocitrate dehydrogenase than to the IDH2 catalytic subunit. After expression in S. cerevisiae from an S. cerevisiae promoter, it was shown to functionally complement an S. cerevisiae idh1 mutant, but not an idh2 mutant, for growth on acetate as a carbon source and for production of NAD(+)‐isocitrate dehydrogenase enzyme activity. These results confirm that the H. capsulatum cDNA encodes a homologue of subunit I of the S. cerevisiae mitochondrial isocitrate dehydrogenase isozyme that functions in the citric acid cycle. The HcIDH1 cDNA sequence is available in GenBank with Accession No. AF009036. Copyright © 1999 John Wiley & Sons, Ltd.

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Subunit structure, expression, and function of NAD(H)-specific isocitrate dehydrogenase in Saccharomyces cerevisiae

Cited by 105 publications

References 40 publications

Purification and Characterization of NAD-Isocitrate Dehydrogenase from Chlamydomonas reinhardtii1

Purification and Characterization of NAD-Isocitrate Dehydrogenase from Chlamydomonas reinhardtii1

Suppression of metabolic defects of yeast isocitrate dehydrogenase and aconitase mutants by loss of citrate synthase

Isolation of aHistoplasma capsulatum cDNA that complements a mitochondrial NAD(+)-isocitrate dehydrogenase subunit I-deficient mutant ofSaccharomyces cerevisiae

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