Molecular Cloning of the Full-length cDNA of (S)-Hydroxynitrile Lyase from Hevea brasiliensis

Hasslacher, Meinhard; Schall, Michael; Hayn, Marianne; Griengl, Herfried; Kohlwein, Sepp D.; Schwab, Helmut

doi:10.1074/jbc.271.10.5884

Cited by 128 publications

(102 citation statements)

References 33 publications

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“…The amino acid sequences of ACC1 of yeast (21), rat (22), chicken (23), and human (12) are very similar, with Ϸ90% identity among the animal ACC1 carboxylases and Ϸ35% similarity between the animal and yeast ACC1 enzymes. In the yeast, Saccharomyces cerevisiae, acetyl-CoA carboxylase (ACC1͞FAS3), the ACC homolog of animal ACC1, is essential for the growth and viability of the yeast cells, and supplementation of fatty acids failed to rescue the acc1 null mutants (24,25).To dissect the roles of the two animal ACC isoforms, we perused knockouts of the ACC1 and ACC2 genes in mice. Our studies with Acc2 knockout mice have shown that Acc2 Ϫ/Ϫ mice live and breed well, continuously oxidize fatty acids, eat more food, and gain less weight than their wild-type cohorts (4).…”

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Mutant mice lacking acetyl-CoA carboxylase 1 are embryonically lethal

Abu-Elheiga

Matzuk²,

Kordari³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

225

162

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Acetyl-CoA carboxylases (ACC1 and ACC2) catalyze the carboxylation of acetyl-CoA to form malonyl-CoA, an intermediate metabolite that plays a pivotal role in the regulation of fatty acid metabolism. We previously reported that ACC2 null mice are viable, and that ACC2 plays an important role in the regulation of fatty acid oxidation through the inhibition of carnitine palmitoyltransferase I, a mitochondrial component of the fatty-acyl shuttle system. Herein, we used gene targeting to knock out the ACC1 gene. The heterozygous mutant mice (Acc1 ؉/؊ ) had normal fertility and lifespans and maintained a similar body weight to that of their wild-type cohorts. The mRNA level of ACC1 in the tissues of Acc1 ؉/؊ mice was half that of the wild type; however, the protein level of ACC1 and the total malonyl-CoA level were similar. In addition, there was no difference in the acetate incorporation into fatty acids nor in the fatty acid oxidation between the hepatocytes of Acc1 ؉/؊ mice and those of the wild type. In contrast to Acc2 ؊/؊ mice, Acc1 ؊/؊ mice were not detected after mating. Timed pregnancies of heterozygotes revealed that Acc ؊/؊ embryos are already undeveloped at embryonic day (E)7.5, they die by E8.5, and are completely resorbed at E11.5. Our previous results of the ACC2 knockout mice and current studies of ACC1 knockout mice further confirm our hypotheses that malonyl-CoA exists in two independent pools, and that ACC1 and ACC2 have distinct roles in fatty acid metabolism. malonyl-CoAA cetyl-CoA carboxylases (ACCs) are biotin-containing enzymes that catalyze the carboxylation of acetyl-CoA to form malonyl-CoA, an intermediate metabolite that plays a key role in the regulation of fatty acid metabolism. In addition, malonylCoA, as a precursor of the synthesis of long-chain fatty acids, has been implicated as a signal molecule for insulin secretion from the pancreatic ␤-islets (1, 2). The role that ACC plays in energy metabolism in lipogenic tissues (liver and adipose) and in oxidative tissues (liver, heart, and skeletal muscle) have become the focus of many studies (3-8). In lipogenic tissues, malonylCoA is the C 2 -unit donor for de novo synthesis of long-chain fatty acids catalyzed by fatty acid synthase (FAS) and for the chain elongation of fatty acid to very long-chain fatty acids. Moreover, increasing evidence suggests that malonyl-CoA is a regulator of fatty acid oxidation through the inhibition of carnitine palmitoyltransferase I, an enzyme that controls the entry of long-chain fatty acids into the mitochondria for ␤-oxidation (3). In animals, including humans, ACC1 (M r ϭ 265,000) and ACC2 (M r ϭ 280,000) are the two isoforms of acetyl-CoA carboxylase that are encoded by two separate genes, ACC1 and ACC2, respectively, and they display distinct tissue distribution (9-12). ACC1 is abundant in lipogenic tissues, such as liver and adipose tissue, whereas ACC2 is highly expressed in heart, skeletal muscle, and liver (9, 10). Recently, Loftus et al. (13) proposed that malonylCoA plays a role in the central ne...

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confidence: 99%

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confidence: 99%

Mutant mice lacking acetyl-CoA carboxylase 1 are embryonically lethal

Abu-Elheiga

Matzuk²,

Kordari³

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

225

162

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“…In yeast, cytoplasmic ACC is a single trifunctional polypeptide of 2233 amino acids and with an approximate molecular mass of 250 kDa (10 -12). The ACC-encoding gene has been isolated and was designated as ACC1 and FAS3, respectively (11,12). Unlike fatty-acid synthase mutants, which grow upon fatty acid supplementation, ACC1 disruption is lethal and cannot be compensated by external long-chain fatty acids (12,13).…”

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“…The ACC-encoding gene has been isolated and was designated as ACC1 and FAS3, respectively (11,12). Unlike fatty-acid synthase mutants, which grow upon fatty acid supplementation, ACC1 disruption is lethal and cannot be compensated by external long-chain fatty acids (12,13). This characteristic is commonly attributed to the malonyl-CoA requirement of cellular very long-chain fatty acid biosynthesis.…”

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“…For many years, however, the biochemical function of HFA1 remained elusive because hfa1⌬ disruptants were, other than acc1 mutants, fatty acid-prototrophic and exhibited no obvious ACC deficiency. Accordingly, the fatty acid-requiring phenotype of ACC1 mutants was not compensated by functional HFA1 DNA (12,13). A first clue to the physiological function of HFA1 came from an observation in our laboratory that HFA1 mutants failed to grow on non-fermentable carbon sources.…”

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HFA1 Encoding an Organelle-specific Acetyl-CoA Carboxylase Controls Mitochondrial Fatty Acid Synthesis in Saccharomyces cerevisiae

Hoja

Marthol

Hofmann

et al. 2004

Journal of Biological Chemistry

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The Saccharomyces cerevisiae gene, HFA1, encodes a >250-kDa protein, which is required for mitochondrial function. Hfa1p exhibits 72% overall sequence similarity (54% identity) to ACC1-encoded yeast cytoplasmic acetyl-CoA carboxylase. Nevertheless, HFA1 and ACC1 functions are not overlapping because mutants of the two genes have different phenotypes and do not complement each other. Whereas ACC1 is involved in cytoplasmic fatty acid synthesis, the phenotype of hfa1⌬ disruptants resembles that of mitochondrial fatty-acid synthase mutants. They fail to grow on lactate or glycerol, and the mitochondrial cofactor, lipoic acid, is reduced to <10% of its normal cellular concentration. Other than Acc1p, the N-terminal sequence of Hfa1p comprises a canonical mitochondrial targeting signal together with a matrix protease cleavage site. Accordingly, the HFA1-encoded protein was specifically assigned by Western blotting of appropriate cell fractions to the mitochondrial compartment. Removal of the mitochondrial targeting sequence abolished the competence of HFA1 DNA to complement hfal null mutants. Conversely and in contrast to the intact HFA1 sequence, the signal sequence-free HFA1 gene complemented the mutational loss of cytoplasmic acetyl-CoA carboxylase. Expression of HFA1 under the control of the ACC1 promoter restored cellular ACC activity in ACC1-defective yeast mutants to wild type levels. From this finding, it is concluded that HFA1 encodes a specific mitochondrial acetyl-CoA carboxylase providing malonyl-CoA for intraorganellar fatty acid and, in particular, lipoic acid synthesis.

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Hydroxynitrile lyase from Hevea brasiliensis: Molecular characterization and mechanism of enzyme catalysis

Hasslacher¹,

Kratky²,

Griengl³

et al. 1997

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(S)-Hydroxynitrile lyase (Hnl) from the tropical rubber tree Hevea brasiliensis is a 29 kDa single chain protein that catalyses the breakdown or formation of a C--C bond by reversible addition of hydrocyanic acid to aldehydes or ketones. The primary sequence of Hnl has no significant homology to known proteins. Detailed homology investigations employing PROFILESEARCH and secondary structure prediction algorithms suggest that Hnl is a member of the alpha/beta hydrolase fold protein family and contains a catalytic triad as functional residues for catalysis. The significance of predicted catalytic residues was tested and confirmed by site-directed mutagenesis and expression of mutant and wild-type proteins in the yeast, Saccharomyces cerevisiae. Based on these data we suggest a mechanistic model for the (S)-cyanohydrin synthesis catalyzed by hydroxynitrile lyase from Hevea brasiliensis.

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Molecular Cloning of the Full-length cDNA of (S)-Hydroxynitrile Lyase from Hevea brasiliensis

Cited by 128 publications

References 33 publications

Mutant mice lacking acetyl-CoA carboxylase 1 are embryonically lethal

Mutant mice lacking acetyl-CoA carboxylase 1 are embryonically lethal

HFA1 Encoding an Organelle-specific Acetyl-CoA Carboxylase Controls Mitochondrial Fatty Acid Synthesis in Saccharomyces cerevisiae

Hydroxynitrile lyase from Hevea brasiliensis: Molecular characterization and mechanism of enzyme catalysis

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