Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.6.1.44 (AGT)
 770 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Kynurenine-glyoxylate aminotransferase, alanine-glyoxylate aminotransferase and serine-pyruvate aminotransferase were co-purified and crystallized as yellow cubes from human liver particulate fraction. The crystalline enzyme was homogeneous by the criteria of electrophoresis, isoelectric focusing, gel filtration, sucrose-density-gradient centrifugation and analytical ultracentrifugation. The molecular weight of the enzyme was calculated as approx. 90000, 89000 and 99000 by the use of gel filtration, analytical ultracentrifugation and sucrose-density-gradient centrifugation respectively, with two identical subunits. The enzyme has a s(20,w) value of 5.23S, an isoelectric point of 8.3 and a pH optimum between 9.0 and 9.5. The enzyme solution showed absorption maxima at 280 and 420nm. The enzyme catalysed transamination between several l-amino acids and pyruvate or glyoxylate. The order of effectiveness of amino acids was alanine>serine>glutamine>glutamate>methionine>kynurenine = phenylalanine = asparagine>valine>histidine>lysine>leucine>isoleucine>arginine>tyrosine = threonine>aspartate, with glyoxylate as amino acceptor. The enzyme was active with glyoxylate, oxaloacetate, hydroxypyruvate, pyruvate, 4-methylthio-2-oxobutyrate and 2-oxobutyrate, but showed little activity with phenylpyruvate, 2-oxoglutarate and 2-oxoadipate, with kynurenine as amino donor. Kynurenine-glyoxylate aminotransferase activity was competitively inhibited by the addition of l-alanine or l-serine. From these results we conclude that kynurenine-glyoxylate aminotransferase, alanine-glyoxylate aminotransferase and serine-pyruvate aminotransferase activities of human liver are catalysed by a single protein. Kinetic parameters for the kynurenine-glyoxylate aminotransferase, alanine-glyoxylate aminotransferase, serine-pyruvate aminotransferase and alanine-hydroxypyruvate aminotransferase reactions of the enzyme are presented.
 ...
PMID:Crystallization and characterization of human liver kynurenine--glyoxylate aminotransferase. Identity with alanine--glyoxylate aminotransferase and serine--pyruvate aminotransferase. 678 36

Alanine-glyoxylate aminotransferase and 2-aminobutyrate aminotransferase were co-purified from rat kidney to a single protein (about 500-fold purified from the homogenate). The activity ratios of alanine-glyoxylate aminotransferase to 2-aminobutyrate aminotransferase were constant during co-purification steps suggesting the 2-aminobutyrate aminotransferase activity was catalysed by only alanine-glyoxylate aminotransferase. The molecular weight of the enzyme was estimated to be approx. 213 000, 220 000 and 236 000 by analytical ultracentrifugation, Sephadex G-150 gel filtration and sucrose density gradient centrifugation, respectively. From the polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate, the enzyme consisted of four apparently similar subunits having a molecular weight of approx. 56 000. The enzyme was almost specific to L-alanine and L-2-aminobutyrate as amino donor and to glyoxylate, pyruvate and 2-oxobutyrate as amino acceptor. The enzyme was identified with rat liver alanine-glyoxylate aminotransferase isoenzyme 2 but not with rat liver alanine-glyoxylate aminotransferase isoenzyme 1 from Ouchterlony double diffusion analysis. Absorption spectra and some kinetic properties of the enzyme were clarified.
 ...
PMID:Co-purification of alanine-glyoxylate aminotransferase with 2-aminobutyrate aminotransferase in rat kidney. 680 44

1. In rats, liver 4-hydroxy-2-oxoglutarate aldolase and hydroxyproline oxidase activities are maximal in the suckling period. 2. Liver activities for 4-hydroxy-2-oxoglutarate aldolase, alanine-glyoxylate aminotransferase, serine-pyruvate aminotransferase and serine dehydratase, but not hydroxyproline oxidase, are increased in rats on a high-fat, carbohydrate-free diet. 3. It is suggested that 4-hydroxy-2-oxoglutarate may be a significant source of glyoxylate for glycine and hence glucose formation. 4. Mammalian liver hydroxyproline oxidase activity is higher in carnivorous species; necessary, perhaps, to metabolise a relatively large influx of hydroxyproline on a flesh diet.
 ...
PMID:Comparative and developmental studies on 4-hydroxy-2-oxoglutarate aldolase and hydroxyproline oxidase. 708 20

Alanine: gamma, delta-dioxovalerate aminotransferase had been purified from bovine liver mitochondria, and the capacity of this enzyme to form delta-aminolevulinic acid had been suggested to be far greater than that of delta-aminolevulinate synthase (EC 2.3.1.37) from the same mitochondria (Varticovski, L., Kushner, J. P., and Burnham, B. F. (1980) J. Biol. Chem. 255, 3742-3747). In the present study, alanine: gamma, delta-dioxovalerate aminotransferase and alanine-glyoxylate aminotransferase (EC 2.6.1.44) were co-purified to homogeneity from bovine liver mitochondria. The ratio of the two activities remains constant during purification and is unchanged by a variety of treatments of the purified enzyme. Alanine: gamma, delta-dioxovalerate aminotransferase activity is competitively inhibited by glyoxylate. Some kinetic data are presented. These results show that the two activities are associated with the same protein. The enzyme is much higher in the glyoxylate aminotransferase activity than in the dioxovalerate aminotransferase activity. The purified enzyme has a molecular weight of approximately 240,000 with four identical subunits and an isoelectric point of 5.4. The ratio of the gamma, delta-dioxovalerate aminotransferase activity to the glyoxylate aminotransferase was determined with alanine:glyoxylate aminotransferase preparations from various mammalian liver and kidney.
 ...
PMID:Biosynthesis of porphyrin precursors in mammals. Identity of alanine: gamma, delta-dioxovalerate aminotransferase with alanine:glyoxylate aminotransferase. 728 16

Rat liver homogenates catalyzed the elimination of fluoride from (R,S)-alpha-fluoro-beta-alanine. The substrate specificity and physical properties of the defluorinating enzyme were similar to those of mitochondrial L-alanine-glyoxylate aminotransferase II (EC 2.6.1.44, AlaAT-II). Furthermore, AlaAT-II activity, measured with L-alanine and glyoxylate as substrates, copurified with the alpha-fluoro-beta-alanine-defluorinating enzyme. The NH2-terminal sequence (18 residues) of the enzyme did not show significant sequence similarity with any of the proteins currently listed in GenBank. The purified enzyme catalyzed the transamination of L-alanine (Ala) and glyoxylate (glyx) at pH 8.5 by a ping-pong mechanism with kinetic parameters of kcat = 17 sec-1, KL-Ala = 3.2 mM, and Kglyx = 0.3 mM, respectively. The kinetic parameters for the defluorination of (R)-alpha-fluoro-beta-alanine and (S)-alpha-fluoro-beta-alanine were kcat = 6.2 and 2.6 min-1, respectively, and Km = 2.7 and 0.88 mM, respectively. L-Alanine potently inhibited the defluorination reaction with an apparent Ki of 0.024 mM. (R,S)-alpha-Fluoro-beta-alanine converted the optical spectrum of the enzyme-bound cofactor from the pyridoxal form to the pyridoxamino form, which indicated that this cofactor may participate in the defluorination reaction. The product of the enzymatic reaction, malonic semialdehyde, reacted nonenzymatically with (R,S)-alpha-fluoro-beta-alanine to form an adduct that was detected spectrally. AlaAT-II was not inactivated during dehalogenation of (R,S)-alpha-fluoro-beta-alanine but was inactivated completely during dehalogenation of beta-chloro-L-alanine.
 ...
PMID:Enzymatic elimination of fluoride from alpha-fluoro-beta-alanine. 750 99

In order to confirm the amino acid sequence predicted from the nucleotide sequence of cDNA and also to elucidate the intracellular localization and molecular evolution, human liver alanine-glyoxylate transaminase 1 (AGT1) was purified and subjected to partial amino acid sequence determination, with special attention to posttranslational modification. The enzyme was purified to homogeneity from the 10,000 x g supernatant of human liver homogenate. The purified enzyme showed only a single protein band at about 43 kDa on SDS-PAGE, indicating that it is a homodimer of two identical subunits, because the native enzyme has a molecular mass of about 80 kDa. Both the amino- and carboxyl-terminal peptides of the enzyme were isolated from a cyanogen bromide digest of the S-carboxyl-methylated protein and subjected to amino acid sequence determination. The alpha-amino group of the amino-terminal peptide was shown to be blocked by an acetyl group. The carboxyl-terminal sequence contained a putative N-glycosylation sequence (-Asn-Ala-Thr-), the only one present in the whole molecule, but this sequence was normally determined, indicating that the enzyme is not N-glycosylated. Purdue et al. [J. Cell Biol. 111, 2341-2351 (1990)] have reported that Pro-11, Gly-170, and Ile-340 in normal human AGT1 were replaced by Leu, Arg, and Met, respectively, in a patient with primary hyperoxaluria type 1. We confirmed that residue-11 was Pro. Both the amino- and carboxyl-terminal sequences of the enzyme showed extensive similarity with those of rat liver mitochondrial serine-pyruvate aminotransferase and the small chain of hydrogenase from a thermophilic unicellular cyanobacterium, Synechococcus PCC 6716.(ABSTRACT TRUNCATED AT 250 WORDS)
 ...
PMID:Purification and amino- and carboxyl-terminal amino acid sequences of alanine-glyoxylate transaminase 1 from human liver. 779 68

1. Cetaben in contrast to fibrates affect differently peroxisomal constituents. 2. Changes in large scale of liver non-peroxisomal parameters were compared after 10 days administration of equal doses (200 mg/kg/day) of cetaben and clofibric acid to male Wistar rats. 3. Clofibric acid treatment increased markedly the activities of FAD-glycerol-3-P dehydrogenase, beta-hydroxyacyl-CoA dehydrogenase, cytochrome-c oxidase, malic enzyme, NAD-glycerol-3-P dehydrogenase, ethoxycoumarin deethylase, p-nitroanisole demethylase and amounts of cytochrome P-450 and b5. 4. However no analogical changes were observed after cetaben treatment in the livers of experimental animals. 5. Both drugs increased the activities of alanine-glyoxylate aminotransferase-1 and acetylcarnitine transferase--enzymes with proven mitochondrial and peroxisomal location. 6. Cetaben contrary to clofibric acid does not increase solubilization of peroxisomal enzymes. 7. Enhanced acetylcarnitine transferase and alanine-glyoxylate aminotransferase-1 activities were distributed in mitochondria as well as in peroxisomes after clofibric acid treatment, however, only peroxisomes were enriched after cetaben administration. 8. The results obtained suggest that cetaben represents an exceptional type of peroxisome proliferator, specifically affecting peroxisomes, without having a negative influence on the processes of peroxisome biogenesis.
 ...
PMID:Cetaben is an exceptional type of peroxisome proliferator. 800 53

In the liver biopsy from an 8.5-year-old girl with the biochemical characteristics of rhizomelic chondrodysplasia punctata (RCDP), but with normal limbs, normal catalase-containing peroxisomes were absent. Light microscopy after diaminobenzidine staining for catalase activity (the peroxisomal marker enzyme) and immunostaining against catalase protein indicated a cytosolic localization of the enzyme. By electron microscopy, rare and extremely large, irregularly shaped vesicles were found in the parenchymal cells. The three peroxisomal beta-oxidation enzymes (acyl-CoA oxidase, bi(tri)functional enzyme, and 3-ketoacyl-CoA thiolase) and alanine-glyoxylate aminotransferase were immunolocalized in these organelles. However, a weak to negative label was obtained after staining against catalase. Diaminobenzidine staining demonstrated a minimal catalase reaction product in some vesicles only. Morphometry revealed a corrected mean d-circle of 1.44 microns and a maximum d-circle of 2.767 microns (controls: 0.635 microns and 1.027 microns, respectively). Numerical, volume, and surface densities were reduced to 3%, 41%, and 17% of control values, respectively. The large size, irregular shape, and rarity of the organelles are morphologic features of peroxisomal "ghosts." It seems that in this patient, apart from the known peroxisomal defects in RCDP, catalase incorporation into the peroxisomes is impaired together with a normal proliferation (division) of the organelles. In the cultured skin fibroblasts from the patient, however, immuno-electron microscopy showed normal catalase-containing peroxisomes in apparently normal numbers.
 ...
PMID:Cytoplasmic catalase and ghostlike peroxisomes in the liver from a child with atypical chondrodysplasia punctata. 812 28

D-3-Aminoisobutyrate-pyruvate aminotransferase (EC 2.6.1.40) and alanine-glyoxylate aminotransferase 2 (EC 2.6.1.44) were co-purified from rat liver as a single protein. The ratio of the two activities remained constant after Sephacryl S-200 chromatography and chromatofocussing. The Km value for beta-alanine as a substrate with 1 mM glyloxylate as amino group acceptor was 1.4 mM. The activity was inhibited by (S)-alanine with Ki = 2.2 mM. The Km for (S)-alanine as substrate with 1 mM glyoxylate as amino group was 6 mM. This activity was inhibited competitively by beta-alanine with Ki = 0.7 mM. (R)-3-aminoisobutyric acid, 5-aminolevulinic acid, NG,NG'-dimethyl-(S)-arginine, and (S)-2-aminobutyric acid were active competitively with respect to beta-alanine with Km of 0.12 mM, 2.1 mM, 6.4 mM and 11.3 mM, respectively. Antiserum to rat liver D-3-aminoisobutyrate-pyruvate aminotransferase inhibited alanine-glyoxylate aminotransferase activity in rat liver in the same way as that of D-3-aminoisobutyrate-pyruvate aminotransferase. Alanine-glyoxylate aminotransferase activity and D-3-aminoisobutyrate-pyruvate aminotransferase activities were inactivated competitively with respect to beta-alanine by 5-fluorouracil and 6-azauracil, which are chemotherapeutic reagents used to cancer. These experiments indicate that D-3-aminoisobutyrate-pyruvate aminotransferase is identical with alanine-glyoxylate aminotransferase 2, aminolevulinate aminotransferase, 2-aminobutyrate aminotransferase and dimetylarginine-pyruvate aminotransferase.
 ...
PMID:Identity of D-3-aminoisobutyrate-pyruvate aminotransferase with alanine-glyoxylate aminotransferase 2. 842 75

The dominant position among oxidoreduction processes in peroxisomes is ascribed to catalase, a number of aerobic oxidases, and Cu,Zn-superoxide dismutase. The peroxidase reaction of catalase requires substrates for hydrogen donation, other than H2O2, e.g. alcohols, aldehydes, formic acid. The peroxisomes contain an alternative system of beta-oxidation of higher carboxylic acids which in some types of plant cells is functionally very closely associated with the glyoxylate cycle. Regarding the role of peroxisomes in the metabolism of carboxylic acids, a very important finding has taken place, namely that besides acyl-CoA synthetase which is specific for long chains, the peroxisomes contain still another enzyme which allows the synthesis of CoA esters of fatty acids with very long chains. It is assumed that the entry of acyl-CoA esters or fatty acids into the perxisomes is performed by means of pores in membranes or acyl-carnitine transferases. Peroxisomes oxidize a very wide scale of substrates and contain several types of acyl-CoA oxidases: palmitoyl-CoA oxidase, pristanoyl-CoA oxidase, trihydroxy-coprostanoyl-CoA oxidase. The second and third reactions of peroxisomal beta-oxidation are catalyzed by the so-called three-functional enzyme, the activities of which are identical to those of 2-enoyl-CoA hydratase, beta-hydroxyacyl-CoA dihydrogenase and enoyl-CoA isomerase. The peroxisomes sufficiently oxidize dicarboxylic acids with a higher number of carbons beginning with the adipic acid. The peroxisomal system of beta-oxidation is utilized in metabolism of prostaglandins, pristanic acid-being the product of phytanic acid alpha-oxidation, and cholesterol. Several enzymatic activities needed for the synthesis of cholesterol partially take place in peroxisomes. The peroxisomes represent a decisive compartment for the initial phases of synthesis of plasmalogens. They contain the following enzymes: NAD(+)-glycerol-P-dehydrogenase, dihydroxyacetone-3-P-acyl-transferase, alkyl-dihydroxyacetone-P synthetase and acyl/alkyl-dihydroxyacetone-P reductase. The metabolism of amino acids takes place under the effect of peroxisomal enzymes--oxidase of diamino acids, D-aspartate oxidase, oxidase of L-pipecolic acid and alanine-glyoxylate aminotransferase. Only a few published sources consider it obvious that liver peroxisomes participate in degradation of spermine and spermidine. Polyamine oxidase oxidizes spermine resulting in the origin of spermidine and 3-aminopropionaldehyde, and spermidine is oxidized to putrescine and 3-aminopropionaldehyde. Peroxisomes in many phylogenetically lower animal species enable the break down of purine bases to urea and glyoxylic acid. In phylogenetically higher primates and in man, the activities of urate oxidase in peroxisomes are absent. (Fig. 14, Ref. 166).
 ...
PMID:The role of peroxisomes in intermediary metabolism. 855 58


<< Previous 1 2 3 4 5 6 7 8 9 Next >>