Isolation and sequence of a cDNA encoding porcine mitochondrial NADP-specific isocitrate dehydrogenase

Haselbeck, Robert J.; Colman, Roberta F.; McAlister-Henn, Lee

doi:10.1021/bi00142a007

Cited by 80 publications

(103 citation statements)

References 25 publications

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“…In contrast, cellular levels of the IDH subunits (IDH1 and IDH2) were substantially elevated (~three-fold relative to parental strain levels, as determined by densitometry) in extracts from the aco1 Δ strain, but were not elevated in extracts from the aco1 Δcit1 Δ strain. Using an antiserum that recognizes both cellular isozymes of NADP + -isocitrate dehydrogenase [18], expression of mitochondrial IDP1 was found to be constitutive in all strains (Fig. 3A), whereas cytosolic IDP2 expression was induced in the idhΔ strain, as mentioned above.…”

Section: Resultssupporting

confidence: 58%

“…The idh1 Δ mutant is not a glutamate auxotroph due to the presence of NADP-specific isocitrate dehydrogenases (IDPs), which are alternate cellular sources of α-ketoglutarate [25]. Mitochondrial IDP1 is constitutively expressed, and expression of cytosolic IDP2 is induced by growth with nonfermentable carbon sources and with galactose ( [10,18] and as described below).…”

Section: Resultsmentioning

confidence: 99%

“…Mark T. McCammon and Gyula Kispal). The antiserum initially prepared using yeast IDP1 also recognizes the IDP2 protein (1:2000 dilution) [18]. Secondary antisera were used at dilutions of 1:5000.…”

Section: Western Blot Analysesmentioning

confidence: 99%

“…The pellets were washed twice with water, then dried for [16][17][18][19][20] h in open tubes placed in a block heater set at 60°C. Dry weights were measured after cooling the tubes for 1-2 h at room temperature; we found that a constant weight was reached using this procedure.…”

Section: Sample Preparation For Metabolite Analysesmentioning

confidence: 99%

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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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Section: Resultssupporting

confidence: 58%

Section: Resultsmentioning

confidence: 99%

Section: Western Blot Analysesmentioning

confidence: 99%

Section: Sample Preparation For Metabolite Analysesmentioning

confidence: 99%

See 2 more Smart Citations

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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“…Lysates from TCA cycle mutants representing each enzyme were probed with antisera to detect Idh1p, Idp2p, and Idp1p, a mitochondrial NADP ϩ -isocitrate dehydrogenase (Haselbeck and McAlister-Henn, 1991). Protein levels of Idh1p were elevated in strains deleted for ACO1 and IDH2 (Figure 7) and appeared to be diminished in several other strains deleted for CIT1, KGD2, and MDH1.…”

Section: Genes Affected By Multiple Tca Cycle Defectsmentioning

confidence: 99%

Global Transcription Analysis of Krebs Tricarboxylic Acid Cycle Mutants Reveals an Alternating Pattern of Gene Expression and Effects on Hypoxic and Oxidative Genes

McCammon

Epstein

Przybyla-Zawislak³

et al. 2003

MBoC

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101

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To understand the many roles of the Krebs tricarboxylic acid (TCA) cycle in cell function, we used DNA microarrays to examine gene expression in response to TCA cycle dysfunction. mRNA was analyzed from yeast strains harboring defects in each of 15 genes that encode subunits of the eight TCA cycle enzymes. The expression of >400 genes changed at least threefold in response to TCA cycle dysfunction. Many genes displayed a common response to TCA cycle dysfunction indicative of a shift away from oxidative metabolism. Another set of genes displayed a pairwise, alternating pattern of expression in response to contiguous TCA cycle enzyme defects: expression was elevated in aconitase and isocitrate dehydrogenase mutants, diminished in α-ketoglutarate dehydrogenase and succinyl-CoA ligase mutants, elevated again in succinate dehydrogenase and fumarase mutants, and diminished again in malate dehydrogenase and citrate synthase mutants. This pattern correlated with previously defined TCA cycle growth–enhancing mutations and suggested a novel metabolic signaling pathway monitoring TCA cycle function. Expression of hypoxic/anaerobic genes was elevated in α-ketoglutarate dehydrogenase mutants, whereas expression of oxidative genes was diminished, consistent with a heme signaling defect caused by inadequate levels of the heme precursor, succinyl-CoA. These studies have revealed extensive responses to changes in TCA cycle function and have uncovered new and unexpected metabolic networks that are wired into the TCA cycle.

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Molecular cloning of the cDNA of mouse mitochondrial NADP-dependent isocitrate dehydrogenase and the expression of the gene during lymphocyte activation

et al. 1996

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The current report documents the molecular cloning of the mouse mitochondrial NADP-dependent isocitrate dehydronegase (mNADP-IDH) cDNA. The cDNA was 1,863 bp in length and contained one open reading frame encoding a 523-residue polypeptide with a predicted molecular weight of 58 kDa. The cDNA and the deduced amino acid (AA) sequence of the mouse mNADP-IDH had a high degree of homology with those of porcine, bovine, alfalfa, and yeast. The recombinant mNADP-IDH expressed in Escherichia coli had active enzymatic function, as well as an expected molecular weight. The heart had the highest constitutive expression of the steady-state mNADP-IDH mRNA, followed by the kidney, while the expression of the gene in other tissues was low. The enzymatic activity of different tissues was in agreement with their mNADP-IDH mRNA levels. The resting lymphocytes had low constitutive expression of the gene, but the steady-state mRNA could be induced 48 h after mitogen stimulation. At the protein level, the resting lymphocytes had low enzymatic activity of mNADP-IDH, but the activity was augmented fivefold after mitogen stimulation. The cytosolic NADP-IDH, on the contrary, remained low or undetectable before and after the mitogen stimulation. Based on our current findings as well as the known roles of the mNADP-IDH in anabolism and in the isocitrate shuttle, it is conceivable that the mNADP-IDH i s necessary for optimizing proliferation in lymphocytes.

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Isolation and sequence of a cDNA encoding porcine mitochondrial NADP-specific isocitrate dehydrogenase

Cited by 80 publications

References 25 publications

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

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

Global Transcription Analysis of Krebs Tricarboxylic Acid Cycle Mutants Reveals an Alternating Pattern of Gene Expression and Effects on Hypoxic and Oxidative Genes

Molecular cloning of the cDNA of mouse mitochondrial NADP-dependent isocitrate dehydrogenase and the expression of the gene during lymphocyte activation

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