Nucleotide sequence for yeast dihydrolipoamide dehydrogenase.

Browning, Karen S.; Uhlinger, David J.; Reed, Lester J.

doi:10.1073/pnas.85.6.1831

Cited by 49 publications

(28 citation statements)

References 17 publications

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“…The near-complete absence in the mutants of overall dehydrogenase activities is due to a parallel decrease in lipoamide dehydrogenase (Table 2), the only component common to both complexes. The possibility that the lipoamide dehydrogenase defect might be caused by mutations in LPD, the structural gene for this enzyme (4,23), was excluded on the basis of the ability of a known lpd mutant to complement the respiratory defect of G178 mutants. The absence of lipoamide dehydrogenase activity was also unlikely to be due to any obvious effect of the mutation on the expression of LPD.…”

Section: Resultsmentioning

confidence: 99%

“…The results of complementation tests made it unlikely that mutations in LPD, the structural gene for lipoamide dehydrogenase (4,23), were responsible for the enzyme deficiency in G178 mutants. The presence of the LPD transcript and the apoprotein in G178 mutants also excluded the possibility of the mutations affecting the expression of the lipoamide dehydrogenase subunit.…”

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confidence: 99%

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Cloning and Characterization of FAD1, the Structural Gene for Flavin Adenine Dinucleotide Synthetase of Saccharomyces cerevisiae

Repetto

Glerum

et al. 1995

Molecular and Cellular Biology

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The FAD1 gene of Saccharomyces cerevisiae has been selected from a genomic library on the basis of its ability to partially correct the respiratory defect of pet mutants previously assigned to complementation group G178. Mutants in this group display a reduced level of flavin adenine dinucleotide (FAD) and an increased level of flavin mononucleotide (FMN) in mitochondria. The restoration of respiratory capability by FAD1 is shown to be due to extragenic suppression. FAD1 codes for an essential yeast protein, since disruption of the gene induces a lethal phenotype. The FAD1 product has been inferred to be yeast FAD synthetase, an enzyme that adenylates FMN to FAD. This conclusion is based on the following evidence. S. cerevisiae transformed with FAD1 on a multicopy plasmid displays an increase in FAD synthetase activity. This is also true when the gene is expressed in Escherichia coli. Lastly, the FAD1 product exhibits low but significant primary sequence similarity to sulfate adenylyltransferase, which catalyzes a transfer reaction analogous to that of FAD synthetase. The lower mitochondrial concentration of FAD in G178 mutants is proposed to be caused by an inefficient exchange of external FAD for internal FMN. This is supported by the absence of FAD synthetase activity in yeast mitochondria and the presence of both extramitochondrial and mitochondrial riboflavin kinase, the preceding enzyme in the biosynthetic pathway. A lesion in mitochondrial import of FAD would account for the higher concentration of mitochondrial FMN in the mutant if the transport is catalyzed by an exchange carrier. The ability of FAD1 to suppress impaired transport of FAD is explained by mislocalization of the synthetase in cells harboring multiple copies of the gene. This mechanism of suppression is supported by the presence of mitochondrial FAD synthetase activity in S. cerevisiae transformed with FAD1 on a high-copy-number plasmid but not in mitochondria of a wild-type strain.Previous biochemical analyses of respiration-defective pet mutants of Saccharomyces cerevisiae (30) led to the identification of two complementation groups lacking ␣-ketoglutarate dehydrogenase (KGDC), as a result of mutations in the KGD1 and KGD2 genes for the dehydrogenase (21) and transsuccinylase (22) components of the complex, respectively. The biochemical screens also disclosed that some complementation groups in the mutant collection are composed of strains deficient in both KGDC and pyruvate dehydrogenase (PDC). Most of the mutants displaying a pleiotropic loss of both activities have been ascertained to be blocked in either the biosynthesis or attachment of lipoic acid to the apoproteins. One complementation group (G178), however, had a distinctly different phenotype. The absence of KGDC and PDC activity in G178 mutants could not be explained by the absence of lipoic acid but, rather, was correlated with a defect in lipoamide dehydrogenase, the only enzyme common to both dehydrogenase complexes (20).The results of complementation tests made it unlikely t...

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Cloning and Characterization of FAD1, the Structural Gene for Flavin Adenine Dinucleotide Synthetase of Saccharomyces cerevisiae

Repetto

Glerum

et al. 1995

Molecular and Cellular Biology

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“…The FLbR was found to be highly related to the family of flavin disulfide oxidoreductases (Williams,199 l), especially the DLDH (EC 1.8.1.4) from various sources. The nucleic acid sequence of FLbR showed identity values ranging from 28 to 56% for the DLDHs from E. coli (Stephens et al, 1983), yeast (Browning et al, 1988), pig (Otulakowski and Robinson, 1987), human (Otulakowski and Robinson, 1987;Pons et al, 1988), and pea (Bourguignon et al, 1992); 18 to 25% for GSHRs (EC 1.6.4.2) from pea (Creissen et al, 1991), human (Krauth-Siegel et al, 1982), and E. coli (Greer and Perham, 1986); 22 to 31% for MERAs (EC 1.16.1.1) from Shigella flexneri (Misra et al, 1985) and Staphylococcus aureus (Laddaga et al, 1987); and 18% for TYTR (EC 1.6.4.8) from…”

Section: Sequence Comparisons Of Flbr With Pyridine Nucleotide-disulfmentioning

confidence: 99%

Cloning and Sequence Analysis of a cDNA Encoding Ferric Leghemoglobin Reductase from Soybean Nodules

Becana

Sarath

et al. 1994

Plant Physiol.

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“…In cell extracts four radioactive protein fractions labeled with D-[2-'4C]riboflayin could be detected after gel filtration, one of which coeluted with lipoamide dehydrogenase activity. The of its procaryotic or eucaryotic origin (4,5,14,18,30,33,50,53,55). The multiplicity of isoenzymes observed from eucaryotic origin was found to be a conformational isomerism (19).…”

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Isolation of an atypically small lipoamide dehydrogenase involved in the glycine decarboxylase complex from Eubacterium acidaminophilum

et al. 1989

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The lipoamide dehydrogenase of the glycine decarboxylase complex was purified to homogeneity (8 U/mg) from cells of the anaerobe Eubacterium acidaminophilum that were grown on glycine. In cell extracts four radioactive protein fractions labeled with D-[2-'4C]riboflayin could be detected after gel filtration, one of which coeluted with lipoamide dehydrogenase activity. The of its procaryotic or eucaryotic origin (4,5,14,18,30,33,50,53,55). The multiplicity of isoenzymes observed from eucaryotic origin was found to be a conformational isomerism (19). Immunological studies suggest that lipoamide dehydrogenase is identical in the pyruvate, 2-oxoglutarate, and branched-chain 2-oxoacid dehydrogenases of the rat heart (26).In enzyme assays pig heart diaphorase is able to replace lipoamide dehydrogenase, protein L, from the chicken liver glycine decarboxylase (12). The lipoamide dehydrogenase of glycine decarboxylase from pea leaf mitochondria exhibits a monomer with an Mr of about 59,000, which is similar to that of the enzyme that is involved in the pyruvate dehydrogenase complex, and monoclonal antibodies raised against lipoamide dehydrogenase inhibit both enzymes to the same extent (51). Therefore, being a conservative enzyme, lipoamide dehydrogenase has also been used to calculate evolutionary relationships (9, 27) to other pyridine nucleotide disulfide oxidoreductase flavoproteins such as glutathione reductase, thioredoxin reductase, and mercuric reductase (11,16,36,54).As the only exception Pseudomonas putida and Pseudomonas aeruginosa produce two lipoamide dehydrogenases during growth on valine, which differ to some degree in molecular masses and kinetic parameters. One is part of 2-oxoglutarate dehydrogenase and probably pyruvate dehydrogenase; the other is part of branched-chain 2-oxoacid dehydrogenase (27,39). During glycine oxidation by P. putida, only the lipoamide dehydrogenase of 2-oxoglutarate dehydrogenase is involved, which has a monomer molecular mass of 56 kilodaltons (kDa), however, rather than the enzyme specific for valine, which has a monomer molecular mass of 49 kDa (38).From all these data it seemed that no specific lipoamide dehydrogenase is involved in glycine oxidation. However, during the isolation of the glycine decarboxylase proteins

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Nucleotide sequence for yeast dihydrolipoamide dehydrogenase.

Abstract: Rabbit antiserum to the dihydrolipoamide dehydrogenase (dihydrolipoamide:NAD' oxidoreductase, EC

Cited by 49 publications

References 17 publications

Cloning and Characterization of FAD1, the Structural Gene for Flavin Adenine Dinucleotide Synthetase of Saccharomyces cerevisiae

Cloning and Characterization of FAD1, the Structural Gene for Flavin Adenine Dinucleotide Synthetase of Saccharomyces cerevisiae

Cloning and Sequence Analysis of a cDNA Encoding Ferric Leghemoglobin Reductase from Soybean Nodules

Isolation of an atypically small lipoamide dehydrogenase involved in the glycine decarboxylase complex from Eubacterium acidaminophilum

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