Carnosine–anserine synthetase of muscle. 4. Partial purification of the enzyme and further studies of β-alanyl peptide synthesis

Stenesh, J.; Winnick, Theodore

doi:10.1042/bj0770575

Cited by 32 publications

(16 citation statements)

References 11 publications

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“…The second one is that recombinant mouse and human ATPGD1 catalyze the ATP-dependent synthesis of carnosine and, with a lower affinity, homocarnosine and other related dipeptides. This rather broad specificity with preference for the formation of carnosine is typical of all preparations of carnosine synthase that have been described until now (11)(12)(13)(14). Furthermore, we confirmed by mass spectrometry the identity of the product made by the recombinant mouse enzyme as carnosine and not the reverse peptide (L-histidyl-␤-alanine), which would have a similar elution profile from ion exchange columns.…”

Section: Discussionsupporting

confidence: 64%

“…Carnosine Synthase Is an ADP-producing Ligase-Our finding that purified chicken carnosine synthase produces stoichiometric amounts of carnosine and ADP indicates that the reac- tion mechanism that had been previously proposed (11,12) has to be revised. This stoichiometric formation of ADP and carnosine excluded the possibility that AMP would be formed and converted to ADP by contaminating adenylate kinase, because 2 molecules of ADP would then be expected to be produced per molecule of carnosine.…”

Section: Discussionmentioning

confidence: 94%

“…Molecular Identification of Carnosine Synthase-The two major advances reported in the present work are the molecular identification of carnosine synthase and the fact that this enzyme catalyzes the hydrolysis of ATP to ADP rather than to AMP as previously assumed (11,12). The two points are linked, because the identification of this enzyme could only be performed once we realized that carnosine synthase converted ATP to ADP and not to AMP, leading to the hypothesis that carnosine synthase belonged to the ATP-grasp family.…”

Section: Discussionmentioning

confidence: 99%

“…Carnosine is known to be synthesized from ␤-alanine and L-histidine by an ATP-dependent synthase (EC 6.3.2.11), which has been partially purified from different sources (11)(12)(13)(14) and shown to also catalyze the synthesis of homocarnosine. The fate of ATP in the reaction has never been directly demonstrated, but based on indirect arguments, the nucleotide triphosphate is thought to be converted to AMP and inorganic pyrophosphate.…”

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confidence: 99%

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Molecular Identification of Carnosine Synthase as ATP-grasp Domain-containing Protein 1 (ATPGD1)

Drożak

Veiga‐da‐Cunha

Vertommen³

et al. 2010

Journal of Biological Chemistry

171

146

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Carnosine (␤-alanyl-L-histidine) and homocarnosine (␥-aminobutyryl-L-histidine) are abundant dipeptides in skeletal muscle and brain of most vertebrates and some invertebrates. The formation of both compounds is catalyzed by carnosine synthase, which is thought to convert ATP to AMP and inorganic pyrophosphate, and whose molecular identity is unknown. In the present work, we have purified carnosine synthase from chicken pectoral muscle about 1500-fold until only two major polypeptides of 100 and 90 kDa were present in the preparation. Mass spectrometry analysis of these polypeptides did not yield any meaningful candidate. Carnosine formation catalyzed by the purified enzyme was accompanied by a stoichiometric formation, not of AMP, but of ADP, suggesting that carnosine synthase belongs to the "ATP-grasp family" of ligases. A data base mining approach identified ATPGD1 as a likely candidate. As this protein was absent from chicken protein data bases, we reconstituted its sequence from a PCR-amplified cDNA and found it to fit with the 100-kDa polypeptide of the chicken carnosine synthase preparation. Mouse and human ATPGD1 were expressed in HEK293T cells, purified to homogeneity, and shown to catalyze the formation of carnosine, as confirmed by mass spectrometry, and of homocarnosine. Specificity studies carried out on all three enzymes were in agreement with published data. In particular, they acted with 15-25-fold higher catalytic efficiencies on ␤-alanine than on ␥-aminobutyrate. The identification of the gene encoding carnosine synthase will help for a better understanding of the biological functions of carnosine and related dipeptides, which still remain largely unknown.

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Section: Discussionsupporting

confidence: 64%

Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Molecular Identification of Carnosine Synthase as ATP-grasp Domain-containing Protein 1 (ATPGD1)

Drożak

Veiga‐da‐Cunha

Vertommen³

et al. 2010

Journal of Biological Chemistry

171

146

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“…Synthesized endogenously by carnosine-anserine synthetase in many eukaryotes, it is found in long-lived mammalian tissues (such as neurons and muscle tissue) at concentrations up to 20 mM (37,41). It has been shown that protein cross-links induced in vitro by methyglyoxal are eliminated in the presence of carnosine (24).…”

Section: Discussionmentioning

confidence: 99%

Antiglycation Effects of Carnosine and Other Compounds on the Long-Term Survival of Escherichia coli

Pepper

Farrell

Nord

et al. 2010

Appl Environ Microbiol

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Glycation, or nonenzymatic glycosylation, is a chemical reaction between reactive carbonyl-containing compounds and biomolecules containing free amino groups. Carbonyl-containing compounds include reducing sugars such as glucose or fructose, carbohydrate-derived compounds such as methylglyoxal and glyoxal, and nonsugars such as polyunsaturated fatty acids. The latter group includes molecules such as proteins, DNA, and amino lipids. Glycation-induced damage to these biomolecules has been shown to be a contributing factor in human disorders such as Alzheimer's disease, atherosclerosis, and cataracts and in diabetic complications. Glycation also affects Escherichia coli under standard laboratory conditions, leading to a decline in bacterial population density and long-term survival. Here we have shown that as E. coli aged in batch culture, the amount of carboxymethyl lysine, an advanced glycation end product, accumulated over time and that this accumulation was affected by the addition of glucose to the culture medium. The addition of excess glucose or methylglyoxal to the culture medium resulted in a dose-dependent loss of cell viability. We have also demonstrated that glyoxylase enzyme GloA plays a role in cell survival during glycation stress. In addition, we have provided evidence that carnosine, folic acid, and aminoguanidine inhibit glycation in prokaryotes. These agents may also prove to be beneficial to eukaryotes since the chemical processes of glycation are similar in these two domains of life.

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Ligases

Spector

1982

Covalent Catalysis by Enzymes

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Carnosine–anserine synthetase of muscle. 4. Partial purification of the enzyme and further studies of β-alanyl peptide synthesis

Cited by 32 publications

References 11 publications

Molecular Identification of Carnosine Synthase as ATP-grasp Domain-containing Protein 1 (ATPGD1)

Molecular Identification of Carnosine Synthase as ATP-grasp Domain-containing Protein 1 (ATPGD1)

Antiglycation Effects of Carnosine and Other Compounds on the Long-Term Survival of Escherichia coli

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