Rat liver dimethylglycine dehydrogenase

Lang, Helmut; Polster, Martin; Brandsch, Roderich

doi:10.1111/j.1432-1033.1991.tb16083.x

Cited by 39 publications

(32 citation statements)

References 31 publications

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“…1). The alanine residue at position 43 of Me 2GlyDH, located N-terminally to the putative mitochondrial processing site [24], is also conserved. The 12-residue sequence corresponding to the SarDH tryptic peptide isolated by Cook et al [9] is located at residues 102Ϫ113 from the total deduced SarDH sequence.…”

Section: Resultsmentioning

confidence: 99%

“…The alignment of the SarDH and Me 2 GlyDH sequences [24] indicated an amino acid similarity spread over the entire polypeptide length with small clusters of conserved residues (Fig. 1).…”

Section: Discussionmentioning

confidence: 99%

“…While the two enzymes are obviously derived from the same ancestor, they seem to have differentiated early in evolution since their degree of similarity is relatively low. The Nterminal dinucleotide-binding motif (Gly74-Xaa-Gly-Xaa-XaaGly79) is conserved [24], while the putative C-terminal H 4 PteGlu 5 binding motif, which is a region rich in positively charged residues [27], shows three substitutions of Lys for Arg. It is noteworthy that the alanine residue located N-terminally to the putative mitochondrial processing site [24] is conserved between SarDH and Me 2 GlyDH.…”

Section: Discussionmentioning

confidence: 99%

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Molecular cloning and tissue distribution of rat sarcosine dehydrogenase

Bergeron

Otto

Blache

et al. 1998

European Journal of Biochemistry

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Sarcosine dehydrogenase (SarDH) is a mitochondrial flavoenzyme involved in the oxidative degradation of choline to glycine. The absence of SarDH activity in humans is genetically transmitted and is the cause of an amino acid metabolism disorder called sarcosinemia. Tryptic fragments of the purified enzyme from rat liver were subjected to Edman degradation and the sequences obtained were used to clone the cDNA encoding the full length protein. The deduced amino acid sequence of SarDH shares an overall similarity of 47% with dimethylglycine dehydrogenase (Me 2 GlyDH), another flavoenzyme involved in the mitochondrial choline catabolism with a similar FAD-binding domain. Covalent binding of FAD to SarDH was demonstrated by the observation of strong fluorescence at 530 nm under excitation at 450 nm of the enzyme immunoprecipitated under denaturing conditions from liver extracts. The localization of SarDH immunoreactivity in the mitochondrial matrix was confirmed by Western-blot analysis of purified mitochondrial fractions. Finally, the tissue distribution of SarDH was investigated by Northern-blot analysis of total RNA and Western-blot analysis of total protein from several rat tissues. A strong expression in the liver, but also in the lung, pancreas, kidney, thymus, and oviduct was observed. We therefore suggest that the enzymes of the choline catabolism pathway are important also for metabolism in nonhepatic tissues.

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Section: Resultsmentioning

confidence: 99%

“…The alignment of the SarDH and Me 2 GlyDH sequences [24] indicated an amino acid similarity spread over the entire polypeptide length with small clusters of conserved residues (Fig. 1).…”

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Molecular cloning and tissue distribution of rat sarcosine dehydrogenase

Bergeron

Otto

Blache

et al. 1998

European Journal of Biochemistry

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“…Lower identities (14.9, 14.6, and 12.3%) were found with the ␤-subunit of the heterotetrameric sarcosine oxidase from Corynebacterium sp. P-1 (49), with the N terminus of dimethylglycine dehydrogenase from rat liver (50), and with the amino acid deaminase from Proteus mirabilis (51). High identities occurred with a not yet identified gene product (accession number U23529) from Caenorhabditis elegans.…”

Section: Purification Of Sarcosinementioning

confidence: 98%

Cloning and Functional Expression of a Mammalian Gene for a Peroxisomal Sarcosine Oxidase

Reuber

Karl

Reimann

et al. 1997

Journal of Biological Chemistry

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Sarcosine oxidation in mammals occurs via a mitochondrial dehydrogenase closely linked to the electron transport chain. An additional H 2 O 2 -producing sarcosine oxidase has now been purified from rabbit kidney. A corresponding cDNA was cloned from rabbit liver and the gene designated sox. This rabbit sox gene encodes a protein of 390 amino acids and a molecular mass of 44 kDa identical to the molecular mass estimated for the purified enzyme. Sequence analysis revealed an N-terminal ADP-␤␣␤-binding fold, a motif highly conserved in tightly bound flavoproteins, and a C-terminal peroxisomal targeting signal 1. Sarcosine oxidase from rabbit liver exhibits high sequence homology (25-28% identity) to monomeric bacterial sarcosine oxidases. Both purified sarcosine oxidase and a recombinant fusion protein synthesized in Escherichia coli contain a covalently bound flavin, metabolize sarcosine, L-pipecolic acid, and L-proline, and cross-react with antibodies raised against L-pipecolic acid oxidase from monkey liver. Subcellular fractionation demonstrated that sarcosine oxidase is a peroxisomal enzyme in rabbit kidney. Transfection of human fibroblast cell lines and CV-1 cells (monkey kidney epithelial cells) with the sox cDNA resulted in a peroxisomal localization of sarcosine oxidase and revealed that the import into the peroxisomes is mediated by the peroxisomal targeting signal 1 pathway.In mammals a variety of H 2 O 2 -producing oxidases including D-amino-acid oxidase, D-aspartate oxidase, L-hydroxy-acid oxidase, acyl-CoA oxidase, and L-pipecolic acid oxidase are compartmentalized in peroxisomes. The H 2 O 2 generated from these reactions is then converted to H 2 O and O 2 by the peroxisomal matrix enzyme catalase (1). Several disorders have been described in which there is a defect in peroxisomal assembly that results in a partial or total absence of peroxisomal functions (for a review see Ref.2). Patients with these peroxisomal disorders such as Zellweger syndrome, neonatal adrenoleukodystrophy, infantile Refsum disease, and hyperpipecolatemia all have elevated levels of L-pipecolic acid, an imino acid, which in human and monkey liver is oxidized by a peroxisomal L-pipecolic acid oxidase (3). Indeed, L-pipecolic acid oxidase activity was not detected in liver samples from patients with Zellweger syndrome (4). Primates dehydrogenate L-pipecolic acid to ␦-piperideine-6-carboxylate which is spontaneously converted to ␣-aminoadipic acid ␥-semialdehyde.The subcellular localization of this pathway seems to differ in other mammalian species. In rabbits, guinea pigs, dogs, and sheep L-pipecolic acid oxidation is primarily mitochondrial (5). However, during our studies examining the subcellular distribution of L-pipecolic acid oxidation in rabbits, a considerable amount of L-pipecolic acid oxidation was detected in the peroxisomes, in addition to the previously reported mitochondrial activity (6). Interestingly, this peroxisomal enzyme showed a high specific activity for sarcosine and also oxidized L-pipecolic acid and L-p...

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“…The former pair of enzymes share 11 identical amino acid residues in a stretch of 15 flanking the 8a-N3-histidyl linkage (mainly C-terminal to the covalent link to flavin), whereas the latter pair have 4 identical amino acids immediately on the N-terminal side of the covalent link to flavin. Other similarities in sequence around the site of attachment are restricted to enzymes that are related throughout the whole of their primary sequences, e.g., tri-and dimethylamine dehydrogenases (Boyd et al, 1992;Yang et al, 1995).It has been observed that several covalent flavoproteins (succinate dehydrogenase, fumarate reductase, dimethylglycine dehydrogenase, monoamine oxidase, sarcosine oxidase) (Reuber et ai., 1997), and flavocytochrome csulfide dehydrogenase (Lang et al, 1991), and the putative covalent flavoprotein fructosyl amino acid oxidase have Gly-X-Gly-X-X-Gly-dinucleotide binding-fold consensus sequences very close to the N-termini, which can be quite distant in the primary structure from the flavin attachment site. This motif is also found in enzymes with noncovalently bound flavin (van Driessche et al, 1996); however, L-gulono-y-lactone oxidase, p-cresol methylhydroxylase, and 6-hydroxy-~-nicotine oxidase are covalent flavoproteins that do not have this so-called "fingerprint" (Brandsch, 1993;Kim et al, 1994).…”

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confidence: 99%

Covalent attachment of flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) to enzymes: The current state of affairs

1998

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The first identified covalent flavoprotein, a component of mammalian succinate dehydrogenase, was reported 42 years ago. Since that time, more than 20 covalent flavoenzymes have been described, each possessing one of five modes of FAD or FMN linkage to protein. Despite the early identification of covalent flavoproteins, the mechanisms of covalent bond formation and the roles of the covalent links are only recently being appreciated. The main focus of this review is, therefore, one of mechanism and function, in addition to surveying the types of linkage observed and the methods employed for their identification. Case studies are presented for a variety of covalent flavoenzymes, from which general findings are beginning to emerge.Keywords: covalent flavoproteins; flavin adenine dinucleotide; flavin mononucleotide; flavinylation; redox cofactors Cofactors are recruited by protein molecules to extend the chemistry available within the active sites of enzymes. The chemistries displayed are variable and the range of protein-specific cofactors available bears testimony to the types of biological reaction achieved by many enzyme molecules. Cofactors may be present in the freely dissociable form, but others, such as lipoic acid and biotin, are permanently linked to the host protein. Other cofactors (e.g., heme and pyridoxyl phosphate) fall into both categories in that they can be attached covalently to the protein or operate in the freely dissociable form. The covalent addition of specific cofactors to unique residues within a polypeptide chain is a fundamental problem in studies of biological molecular recognition. In many cases, this specificity is purchased at the expense of recruiting modification enzymes that direct the covalent addition of a cofactor to the host protein. For example, the additions of biotin and lipoic acid to the €-amino groups of specific Lys residues in carboxyltransferases and the 2-oxoacid dehydrogenases are directed by a biotin holoenzyme synthase (Sweetman et al., 1985;Takai et al., 1987) and Reprint requests to: N.S. Scmtton, Department of Biochemistry, University of Leicester, Adrian Building, University Road, Leicester LE1 7RH UK; e-mail: nss4@le.ac.uk; or W.S. McIntire, Molecular Biology Division, Department of Veterans Affairs Medical Center, San Francisco, California 94121; e-mail: wsm@itsa.ucsf.edu.lipoate-protein ligases (Brookfield et al., 1991; Moms et al., 1994), respectively. In a similar fashion, a heme cytochrome c lyase (Steiner et al., 1996) is responsible for the covalent addition of heme to two Cys residues via thioether linkages. In some enzymes, an existing side chain is modified to yield what is in effect a covalently attached cofactor, although no pre-existing cofactor molecule is incorporated into the enzyme. Several examples of this type of modification are provided in the next paragraph.For histidine decarboxylase of Lactobacillus 30A (Recsei & Snell, 1984), a pyruvoyl group is generated from an existing residue, Ser 82. The enzyme is synthesized as a proenzym...

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Rat liver dimethylglycine dehydrogenase

Cited by 39 publications

References 31 publications

Molecular cloning and tissue distribution of rat sarcosine dehydrogenase

Molecular cloning and tissue distribution of rat sarcosine dehydrogenase

Cloning and Functional Expression of a Mammalian Gene for a Peroxisomal Sarcosine Oxidase

Covalent attachment of flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) to enzymes: The current state of affairs

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