EC Number   |
Recommended Name |
Application |
|---|
   4.6.1.13 | phosphatidylinositol diacylglycerol-lyase |
analysis |
use of the enzyme and specific antibodies for the enzyme for the examination of the growth inhibition, morphological change and ectoenzyme release of the LLC-PK1 cells from pig, effective for the investigation of the function of the glycosyl-phosphatidylinositol-anchor protein |
| 4.6.1.18 | pancreatic ribonuclease |
analysis |
preparation of polymeric nanoparticles imprinted with RNase A via miniemulsion polymerization using methyl methacrylate and ethylene glycol dimethacrylate. The addition of poly(vinyl alcohol) as a co-surfactant is effective in preserving the protein structural integrity. Imprinted nanoparticles produces by the optimized method show increased target specificity |
| 4.6.1.18 | pancreatic ribonuclease |
analysis |
pressure tuning hole burning experiments using the UV-absorbing tyrosine residues. Ribonuclease A protein stays intact upon cooling to 2 K. Its various tyrosine sites show characteristic features which can be resolved in pressure tuning hole burning spectra. Reducing the sulfur bridges leads to a loss of the individual features, and the sites become alike. The respective compressibility is reduced by more than a factor of 2 and comes close to the value of free tyrosine in solution. Compared to the reduction of the sulfur bridges, the influence of guanidinium hydrochloride on the pressure tuning behavior is less pronounced |
| 4.6.1.18 | pancreatic ribonuclease |
analysis |
studies on application to purify ribonuclease from eggs of Rana catesbeiana with an aqueousaqueous polymer phase system by using a small-scale cross-axis coil planet centrifuge |
| 4.6.1.18 | pancreatic ribonuclease |
analysis |
study on spermidine modulation of enzyme activity via individual RNA plasmon rulers which combine high throughput with high temporal resolution at the single molecule level and are able to retrieve otherwise obscured information about weak structural stabilizations |
| 4.6.1.18 | pancreatic ribonuclease |
analysis |
synthesis of molecularly imprinted polymers from the monomers styren and polyethyleneglycol 400 dimethacylate with high rebinding efficiency of RNase A to polymer. Polymers show high selectivity for RNase A and high stability |
| 4.6.1.18 | pancreatic ribonuclease |
analysis |
synthesis of monodispersed RNase A surface-imprinted particles with good magnetic property for practical bioseparation. Use of methyl methacrylate and ethylene glycol dimethacrylate as the functional and cross-linker monomers to produce particles of 700-800 nm diameter imprinted with ribonuclease A and encapsulated with nanosized Fe3O4 particles. Surface-imprinted particles show good selectivity toward the RNase template over control protein |
| 4.6.1.20 | ribonuclease U2 |
analysis |
bacteriophage lambda protein phosphatase and recombinant RNase U2 can be used to generate RNase digestion products amenable to liquid chromatography-tandem mass spectrometry analysis, which can be used to determine the presence and location of modified ribonucleosides in RNA samples |
| 4.6.1.20 | ribonuclease U2 |
analysis |
improvement of RNA modification mapping sequence coverage by LC-MS. The non-specific ribonuclease activity RNase U2 E49A substitution mutant provides increased sequence coverage of substrate RNA during modification mapping. Data analysis software is modified to account for non-specific digestion. This combination allows efficient and accurate RNA modification mapping |
| 4.6.1.21 | Enterobacter ribonuclease |
analysis |
- |
 4.98.1.1 | protoporphyrin ferrochelatase |
analysis |
Pro255 has a crucial role in maintaining an appropriate protein conformation and modulating the selectivity and/or regiospecificity of ferrochelatase, ferrochelatase mutants with improved tolerance towards N-methylprotoporphyrin may be potentially used in cell assay systems to study physiological responses to haem deficiency |
| 4.99.1.8 | heme ligase |
analysis |
development of a high resolution correlative combination of cryo soft X-ray tomography to obtain 3D parasite ultrastructure with cryo X-ray fluorescence microscopy to measure heme concentrations |
| 5.1.1.7 | diaminopimelate epimerase |
analysis |
simple and sensitive spectrophotometric method for the determination of meso-alpha,epsilon-diaminopimelate with meso-2,6-diaminopimelate D-dehydrogenase and its application to the assay of diaminopimelate epimerase |
| 5.1.1.7 | diaminopimelate epimerase |
analysis |
a high-performance liquid chromatography method for the simultaneous assay of diaminopimelate epimerase and decarboxylase |
| 5.1.1.8 | 4-hydroxyproline epimerase |
analysis |
spectrophotometric assay for hydroxyproline in collagenous tissue hydrolysates, with an enzymatic method using 4-hydroxyproline 2-epimerase,EC 5.1.1.8, D-amino acid oxidase, EC 1.4.3.3, and a colorimetric reagent of the mixture of Ti(IV) and 4-(2-pyridylazo)-resorcinol |
| 5.1.1.8 | 4-hydroxyproline epimerase |
analysis |
estimation of the content of L-hydroxyprolines using coupling systems with metabolic enzymes of the trans-4-hydroxy-L-proline pathway (hydroxyproline 2-epimerase (HypE) and cis-4-hydroxy-D-proline dehydrogenase (HypDH)) and the trans-3-hydroxy-L-proline pathway (trans-3-hydroxy-L-proline dehydratase (T3LHypD) and DELTA1-pyrroline-2-carboxylate reductase (Pyr2CR)) from microorganisms. A functional expression system of recombinant HypDH with a heterooligomeric structure is constructed in Escherichia coli cells. Enzymological characterization reveals that the beta-subunit acts as a catalytic subunit, and also that assembly with other subunit(s) improves the kinetics for cis-4-hydroxy-D-proline and thermostability. By using a spectrophotometric assay with different wavelengths, the contents of trans-4-hydroxy-L-proline and trans-3-hydroxy-L-proline are successfully estimated within the ranges of 0.004-1 mM and 0.05-1 mM, respectively, and are consistent with the contents determined by HPLC. This enzymatic method is used to measure the content of trans-4-hydroxy-L-proline in the acid-hydrolysate of collagen, and blood plasma |
| 5.1.1.10 | amino-acid racemase |
analysis |
simple procedure for in situ analysis of stereospecificity of C-4 hydrogen transfer of NADH by an NAD-dependent dehydrogenase by combination with amino acid racemase, EC 5.1.1.10, and L-leucine dehydrogenase, EC 1.4.1.9 |
| 5.1.1.13 | aspartate racemase |
analysis |
estimation of age from dentin by using the racemization reaction of Asp |
| 5.1.2.2 | mandelate racemase |
analysis |
direct kinetic assay for mandelate racemase using circular dichroic measurement |
| 5.1.3.1 | ribulose-phosphate 3-epimerase |
analysis |
spectrophotometric assay for D-ribulose-5-phosphate 3-epimerase and D-ribose-5-phosphate ketol-isomerase |
| 5.1.3.1 | ribulose-phosphate 3-epimerase |
analysis |
direct assay procedure which exploits differences in CD spectrum between ribulose 5-phosphate and xylulose 5-phosphate |
| 5.1.3.1 | ribulose-phosphate 3-epimerase |
analysis |
spectrophotometric determination of ribulose 5-phosphate 3-epimerase in tissue extract |
| 5.1.3.3 | Aldose 1-epimerase |
analysis |
kinetic assay method for aldose 1-epimerase based upon fast in situ generation of alpha-D-glucose employing hydrolysis of sucrose by beta-fructofuranosidase and a subsequent reporter reaction involving the aerobic oxidation of beta-D-glucose via glucose o |
| 5.1.3.3 | Aldose 1-epimerase |
analysis |
rapid determination of glucose by aldose 1-epimerase |
| 5.1.3.3 | Aldose 1-epimerase |
analysis |
conductometric biosensor for sucrose determination using a membrane containing enzymes invertase, mutarotase, and glucose oxidase as a sensitive element immobilized on the conductometric interdigitated planar electrodes. The biosensor demonstrates high selectivity, operational stability and reproducibility |
| 5.1.3.17 | heparosan-N-sulfate-glucuronate 5-epimerase |
analysis |
assay procedure for heparosan-N-sulfate-glucuronate 5-epimerase, which is based on the use of a two-phase system for liquid scintillation counting |
| 5.1.3.19 | chondroitin-glucuronate 5-epimerase |
analysis |
assay procedure for chondroitin-glucuronate 5-epimerase which is based on the use of a two-phase system for liquid scintillation. 3H2O formed during the reaction, is extracted into an organic phase containing fluorine and isoamyl alcohol, while unreacted pol |
| 5.1.3.37 | mannuronan 5-epimerase |
analysis |
development of a high-throughput screening method to discriminate between different alginate structures |
| 5.1.3.37 | mannuronan 5-epimerase |
analysis |
in vitro assay for mannuronan C5 epimerization wherein extracts of Escherichia coli expressing high levels of AlgG are incubated with polymannuronate. Epimerization of D-mannuronate to L-guluronate residues in the polymer is detected enzymatically, using a L-guluronate-specific alginate lyase of Klebsieila aerogenes |
| 5.1.3.37 | mannuronan 5-epimerase |
analysis |
rapid and reproducible assay for the analysis of epimerase activity in the fermentation broth using [3H]-alginate as a substrate. The effects of potential interfering compounds are characterized and the uncertainty of the analysis system has been determined. A standard method suitable for use directly on the fermentation broth with a standard deviation within a single analysis of 2.5% has been developed |
| 5.1.99.1 | methylmalonyl-CoA epimerase |
analysis |
a simple direct assay for DL-methylmalonyl-coenzyme A racemase which is based on the fact that the proton on C-2 of methylmalonyl-CoA is replaced by a proton in the medium during racemization |
| 5.1.99.4 | alpha-methylacyl-CoA racemase |
analysis |
fluorescent enzyme-linked aptamer assay for alpha-methylacyl-CoA racemase. The assay shows a dynamic range from 0.1 to 1000 nM of AMACR, a low detection limit of 0.44 nM (19.5 ng/ml), and high AMACR specificity |
| 5.2.1.8 | peptidylprolyl isomerase |
analysis |
the enzyme weakly catalyzes some protein processes which are rate-limited by proline isomerization, but probably exhibits no measurable catalysis towards others. This somewhat limits the usefulness of the enzyme as a diagnostic reagent for proline isomeri |
| 5.2.1.8 | peptidylprolyl isomerase |
analysis |
high-throughput screening method for isoform FKBP12 binding based on the decrease of a fluorescence signal generated by a small molecule fluorescent FKBP12 ligand bound to the protein and measured in the presence of inhibitor |
| 5.2.1.8 | peptidylprolyl isomerase |
analysis |
fluorescent probe 6-(dimethylamino)-2-naphthoyl-Ala-Ala-(cis)-Pro-Phe-4-nitroanilide can be used for imaging active peptidyl-prolyl cis/trans isomerases in live cells |
| 5.3.1.1 | triose-phosphate isomerase |
analysis |
specific polymerase chain reaction protocols used to determine the prevalence of toxigenic Clostridium difficile in Vhembe, South Africa. The study confirms the usefulness of PCR methodologies in the detection of toxigenic Clostridium difficile and suggests that Clostridium difficile is responsible for a small, but underappreciated, proportion of diarrheal cases in the region |
| 5.3.1.1 | triose-phosphate isomerase |
analysis |
systematic mutagenesis method to find critical residues for certain physico-chemical properties of a protein. The method is a shorter alternative to random mutagenesis, saturation mutagenesis or directed evolution to find multiple amino acids critical for certain properties of proteins |
| 5.3.1.8 | mannose-6-phosphate isomerase |
analysis |
development and optimization of a transformation method using the phosphomannose-isomerase gene pmi as a selectable marker for Brassica napus transformation via Agrobacterium tumefaciens, overview |
| 5.3.1.9 | glucose-6-phosphate isomerase |
analysis |
Pgi is an adaptive marker, rather than providing insights into individual genetic health, Pgi appears to have a role in conservation genetics by providing insights into gene by environment interactions, local adaptation and evolutionary significant units, and potentially even morphologically cryptic dispersal phenotypes, method development, overview |
| 5.3.1.9 | glucose-6-phosphate isomerase |
analysis |
phosphoglucose isomerase variability of Cerastoderma glaucum as a model for testing the influence of environmental conditions and dispersal patterns through quantitative ecology approaches, overview. The PGI locus is an appropriate marker for revealing the spatial distribution of genetic variation and for establishing the relationships between allelic composition and the environmental influencing variables |
| 5.3.1.9 | glucose-6-phosphate isomerase |
analysis |
development of a bioautographic assay based on thin layer chromatography for inhibition of PDI by phosphoenolpyruvate. The detection limit for phosphoenolpyruvate as an inhibitor of PGI is 226 microg per spot/zone |
| 5.3.3.7 | aconitate DELTA-isomerase |
analysis |
a simple sensitive and specific method for measuring aconitate isomerase in plants, which depends on the release of tritium from labeled trans-aconitate |
| 5.3.3.7 | aconitate DELTA-isomerase |
analysis |
method for estimation of enzyme activity in plant material |
| 5.3.4.1 | protein disulfide-isomerase |
analysis |
development of a method to determine quantitatively the redox state of active-site cysteines found in the Cys-Xaa-Xaa-Cys motif in living cells. Method is based on the alkylation of cysteines by methoxy polyethylene glycol 5000 maleimide. In vivo, protein disulfide isomerase is present in two semi-oxidized forms in which either the first active site in the a domain or the second active site in the a' domain is oxidized. In HEK-293 cells, about 50% of enzyme is fully reduced, in 18% a domain is oxidized, a' reduced, in 15%, the a domain is reduced, a' oxidized, and 16% of enzyme are fully oxidized |
| 5.3.4.1 | protein disulfide-isomerase |
analysis |
study on the critical influence of reference genes used for data normalization, shown for protein disulfide isomerase |
| 5.3.4.1 | protein disulfide-isomerase |
analysis |
PDIA3 and C/EBP? may be valuable markers in fish for exposure and effect to environmental stress |
| 5.3.99.2 | Prostaglandin-D synthase |
analysis |
particle concentration fluorescence immunoassay for prostaglandin D synthase is suitable for determining the content of prostaglandin D synthetase in various regions of the rat CNS. The method allows to assay a large number of samples with reasonable sensitivity |
| 5.3.99.2 | Prostaglandin-D synthase |
analysis |
PGD synthase is a useful marker for identifying the differentiation stage of human megakaryocytic cells |
| 5.3.99.5 | thromboxane-A synthase |
analysis |
application of monoclonal antibodies to develop a tandem immunoradiometric assay |
| 5.3.99.5 | thromboxane-A synthase |
analysis |
immobilized enzyme is sufficiently stable to be used as a model for studying the properties of the enzyme |
| 5.4.2.6 | beta-Phosphoglucomutase |
analysis |
a specific method for the quantitative determination of beta-glucose 1-phosphate |
| 5.4.2.7 | phosphopentomutase |
analysis |
a coupled optical enzyme assay |
| 5.4.3.7 | leucine 2,3-aminomutase |
analysis |
separation by high-performance liquid chromatography of alpha-amino acids and beta-amino acids and the application to the assay of leucine 2,3-aminomutase |
| 5.4.99.2 | methylmalonyl-CoA mutase |
analysis |
method for separation of methylmalonyl-CoA and succinyl-CoA by capillary electrophoresis suitable for evaluation of total and holo-enzyme activity in biological matrices. Application of method for the differential diagnosis of methylmalonic acidemia, in relation to protein or coenzyme defects |
| 5.4.99.2 | methylmalonyl-CoA mutase |
analysis |
establishment of a method to measure the concentration of succinyl-CoA with UPLC-MS/MS after enzyme reaction using peripheral lymphocytes, and investigation of the MCM enzyme activity of patients with methylmalonic acidemia |
| 5.4.99.5 | chorismate mutase |
analysis |
As an intramolecular reaction that appears to be catalyzed without intermediate steps, covalent catalysis, or modification of the reaction pathway, the chorismate-prephenate rearrangement has become an important model system for theoretical approaches to the study of enzyme catalysis |
| 5.4.99.9 | UDP-galactopyranose mutase |
analysis |
high-throughput fluorescence polarization assay for indentification of inhibitors of enzyme and homologues |
| 5.5.1.6 | chalcone isomerase |
analysis |
improvements in assay procedure |
| 5.6.2.1 | DNA topoisomerase |
analysis |
the absence of cysteine residues in MtTOP1 makes it an attractive system for introduction of potentially informative chemical or spectroscopic probes at specific positions via cysteine mutagenesis. Such probes could be useful for development of high throughput screening assays and can be utilized for fluorescence probe incorporation and fluorescence resonance energy transfer measurement with fluorophore-labeled oligonucleotide substrate |
| 5.6.2.2 | DNA topoisomerase (ATP-hydrolysing) |
analysis |
use of triple-helical DNA structures for targeting enzyme-mediated cleavage to DNA specific sequences and use of drug-triplex-forming oligonucleotides to investgate enzyme mechanism |
| 6.1.1.21 | histidine-tRNA ligase |
analysis |
alternating catalysis requires a mechanism for coupling events between active sites, presumably through conformational changes propagated between these active sites, a version of HisRS is developed that features the site-specific incorporation of extrinsic environmentally sensitive fluorescent probes, allowing the adenylation reaction to be followed by stopped-flow fluorometry |
| 6.2.1.1 | acetate-CoA ligase |
analysis |
colorimetric assay method to measure acetyl-CoA synthetase activity |
| 6.2.1.7 | cholate-CoA ligase |
analysis |
a coupled assay for bile acid:CoA ligase and glycination of bile acid-CoA catalyzed by bile acid-CoA:glycine N-acetyltransferase |
| 6.2.1.7 | cholate-CoA ligase |
analysis |
the ligase assay can be used as a sensitive enzymic marker for endoplasmic reticulum in rat liver |
| 6.2.1.11 | biotin-CoA ligase |
analysis |
enzymatic assay for biotin |
| 6.2.1.17 | propionate-CoA ligase |
analysis |
rapid radiochemical assay method with high sensitivity and specificity |
| 6.2.1.30 | phenylacetate-CoA ligase |
analysis |
determination of of microbody-borne enzymes by analysis of putative microbody targeting singals and a proteomics based inventory of Penicillium chrysogenum microbody matrix proteins using nano-LC-MS/MS analysis |
| 6.3.1.2 | glutamine synthetase |
analysis |
immunoassay that detects synthetase protein samples where the enzyme has been inactivated by repeated cycles of freezing and thawing, and in serum and cerebrospinal fluid where glutamine synthetase is undetectable by the enzyme activity assay |
| 6.3.1.5 | NAD+ synthase |
analysis |
the enzyme is used for an enzymatic cycling method for ammonia assay using a system consisting of three enzymes: EC 6.3.1.5, EC 1.1.1.47 and EC 1.6.99.2 |
| 6.3.1.5 | NAD+ synthase |
analysis |
method for measurement of allantoin in human serum. Serum allantoin is converted to allantoate by the action of allantoinase,and endogenous ammonia is simultaneously removed by the action of glutamine synthetase II. In the second step, L-methionine sulfoximine is used to inhibit glutamine synthetase II, and ammonia is liberated from allantoate by the activity of allantoate amidohydrolase. The ammonia is then converted to NAD by NAD synthetase. Subsequent action of glucose dehydrogenase and diaphorase acts to cycle the formed NAD between its oxidized and reduced forms, resulting in the production of WST-1 formazan, which is monitored at 450 nm. The assay standard curve is linear from 0 to 70 microM allantoin |
| 6.3.1.5 | NAD+ synthase |
analysis |
use of substrate 2-fluro-ATP as tool for 18F NMR-based activity screening |
| 6.3.2.3 | glutathione synthase |
analysis |
assay of glutathione synthetase in erythrocytes by HPLC with fluorimetric detection, useful for rapid screening of erythrocytes glutathione synthetase activity in various pathological conditions |
| 6.3.2.4 | D-Alanine-D-alanine ligase |
analysis |
assay method for screening for effectors of alanine racemase and/or D-alanine:D-alanine ligase |
| 6.3.2.9 | UDP-N-acetylmuramoyl-L-alanine-D-glutamate ligase |
analysis |
assay for monitoring enzyme activity based on the accumulation of adenosine 5'-diphosphate, a product of the reaction catalyzed by MurD ligase, by conversion to a fluorescent signal via a coupled enzyme system, with counterscreen assay to eliminate false positive results |
| 6.3.2.12 | dihydrofolate synthase |
analysis |
a simple radioassay for dihydrofolate synthetase activity and its application to an inhibition study of new pteroate analogs |
| 6.3.2.17 | tetrahydrofolate synthase |
analysis |
assay procedure for the measurement of FPGS in up to 50 or 100 samples of partially purified enzyme at a time |
| 6.3.2.25 | tubulin-tyrosine ligase |
analysis |
utilization of ligase to determine the state of tyrosination of tubulin |
| 6.3.3.2 | 5-formyltetrahydrofolate cyclo-ligase |
analysis |
development of a functional complementation assay for 5-CHO-THF metabolism in Escherichia coli, based on deleting the gene encoding 5-FCL. The deletion mutant accumulates 5-formyltetrahydrofolate and,with glycine as sole nitrogen source, shows a growth defect, both phenotypes are complemented by bacterial or archaeal genes encoding glutamate formiminotransferase. Glutamate formiminotransferases functionally replace 5-formyltetrahydrofolate cyclo-ligases in certain prokaryotes |
| 6.3.4.2 | CTP synthase (glutamine hydrolysing) |
analysis |
fast assay allows the processing of a large number of samples |
| 6.3.4.6 | urea carboxylase |
analysis |
enzymatic assay with urea amidolyase for determination of potassium in serum |
| 6.3.4.10 | biotin-[propionyl-CoA-carboxylase (ATP-hydrolysing)] ligase |
analysis |
96-well plate assay for high-throughput analysis of holocarboxylase synthetase activity by biotinylation of the polypeptide p67, which comprises the 67 C-terminal amino acids in human propionyl-CoA carboxylase, including the biotin-binding site lysine-669, using IRDye-streptavidin and infrared spectroscopy. The minimal concentration of recombinant HCS that can be detected by this assay is less than 1.08 nmol/l. Jurkat cells contain 0.14 U of HCS activity [in micromol of biotinylated p67 formed/(nmol/l HCS h)] in 400 microg of total protein |
| 6.3.4.10 | biotin-[propionyl-CoA-carboxylase (ATP-hydrolysing)] ligase |
analysis |
fusion of full-length HCS to DNA adenine methyltransferase Dam and subsequent transfection into breast cancer cells MCF-7 and normal breast cells MCF-10A for identification of chromatin binding sites |
| 6.3.4.12 | glutamate-methylamine ligase |
analysis |
assay for the enzymatic synthesis of gamma-glutamylmethylamide |
| 6.3.4.15 | biotin-[biotin carboxyl-carrier protein] ligase |
analysis |
biotin-mediated ATP-diphosphate assay which does not require the isolation of the apoenzyme and is simple and convenient for use on a routine assay. The procedure is specifically designed for assay of enzyme from adipose tissue of biotin-deficient rats. W |
| 6.3.4.15 | biotin-[biotin carboxyl-carrier protein] ligase |
analysis |
biotinylation of apo-carboxyl carrier protein, a subunit of acetyl-CoA carboxylase from E. coli is a sensitive and convenient assay method for biotin-[acetyl-CoA-carboxylase] ligase |
| 6.3.4.15 | biotin-[biotin carboxyl-carrier protein] ligase |
analysis |
SH2 domain-based tyrosine kinase assay using biotin ligase modified with a terbium(III) complex. An SH2 domain from lymphocyte-specific tyrosine kinase is genetically fused to a truncated biotin carboxyl carrier protein, and the resulting fusion protein is labeled through biotinylation with biotin protein ligase carrying multiple copies of a luminescent Tb3+ complex. The labeled SH2 fusion proteins are employed to detect a phosphorylated peptide immobilized on the surface of the microtiter plate, where the phosphorylated peptide is produced by phosphorylation to the substrate peptide by Src tyrosine kinase. The assay allows for a reliable determination of the activity of Src kinase lower than 10 ng/mL |
| 6.3.4.15 | biotin-[biotin carboxyl-carrier protein] ligase |
analysis |
development of a high-throughput assay building on the principle of differential scanning fluorimetry of GFP-tagged proteins |
| 6.3.4.15 | biotin-[biotin carboxyl-carrier protein] ligase |
analysis |
fluorescent probe (4S)-4-[5-(1-[3-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]propyl]-1H-1,2,3-triazol-4-yl)pentyl]tetrahydro-1H-thieno[3,4-d]imidazol-2(3H)-one is synthesized by Bpl in vivo, accumulates in cytoplasm and can be used to gain insights into the mechanism of uptake, efflux and metabolism of BPL inhibitors |
| 6.3.4.16 | carbamoyl-phosphate synthase (ammonia) |
analysis |
development of a CPS1-reporter system for the assessment of ammonia metabolism. Labeling of CPS1 gene in cell lines HepG2, and LO2, with fluorescence protein. Cellular detoxification enhancers are selected among a collection of 182 small molecules. In both CPS1 reporter cell lines, the fluorescence intensity is positively correlated with cellular CPS1 mRNA expression, ammonia elimination and secreted urea, and reflects ammonia detoxification in a dose-dependent manner |
| 6.3.4.19 | tRNAIle-lysidine synthase |
analysis |
development of a ultrahigh-throughput, fluorescence anisotropy-based assay for the incorporation of lysine into Ile2 tRNA |
| 6.3.5.4 | asparagine synthase (glutamine-hydrolysing) |
analysis |
high-performance liquid chromatography assay for Asn synthetase is an extremly sensitive and reliable method for assaying Asn synthetase |
| 6.3.5.4 | asparagine synthase (glutamine-hydrolysing) |
analysis |
a rapid, inexpensive micro assay that can be adapted for large numbers of samples |
| 6.3.5.4 | asparagine synthase (glutamine-hydrolysing) |
analysis |
mass spectrometry-based procedure for the direct quantification of asparagine synthetase protein concentration in complex sample mixtures. Assay is able to distinguish samples from transformed cell lines that express the enzyme over a wide dynamic range of concentration. The method directly detects asparagine synthetase protein, use in blast samples from patients with acute lymphoblastic leukemia |
| 6.3.5.4 | asparagine synthase (glutamine-hydrolysing) |
analysis |
development of a sensitive, non-radioactive assay for AsnS, based on incubation of desalted enzyme and substrates and then direct detection of either product asparagine or glutamate by HPLC |
| 6.3.5.7 | glutaminyl-tRNA synthase (glutamine-hydrolysing) |
analysis |
simple system for monitoring the inhibition of GatCAB activity using Escherichia coli Top10 co-expressing the non-discriminating glutamyl-tRNA synthetase ndGluRS and GatCAB genes from Staphylococcus aureus Mu50. Growth repression is confirmed by introducing ndgluRS from Staphylococcus aureus Mu50 into Escherichia coli. Co-expression of the gatCAB operon alleviates growth repression in the host Escherichia coli. The screening system consists of these two transformants and non-expressing Escherichia coli Top10. The transformant harbors both ndGluRS gene and GatCAB operon can be co-expressed in the presence and in the absence of chemical compounds of interest. There is no inhibitor that inactivates GatCAB activity, but upon expression of two inactive GatCAB deletion variants, GatCAB-10 and GatCAB-CHD, together with ndGluRS in Escherichia coli Top10, the cells show repressed growth as well as ndGluRS is expressed |
| 6.4.1.3 | propionyl-CoA carboxylase |
analysis |
method to analyze propionyl-CoA carboxylase activity in phytohemagglutinin stimulated lymphocytes using high performance liquid chromatography. Propionyl-CoA carboxylase activity is unaffected even when lymphocytes are isolated and phytohemagglutinin stimulated after a whole blood sample has been stored at 4°C for 5 days, and the method is useful for the confirmation of propionic acidemia in individuals, and prenatal diagnosis and genetic counseling for the affected families |
| 6.4.1.8 | acetophenone carboxylase |
analysis |
construction of a reporter strain by inserting a gene fusion of apcA, the first gene of the apc-bal operon, with the gene for the fluorescent protein mCherry into the chromosome. mCherry production is proportional to the applied acetophenone concentrations. The reporter strain allows quantification of acetophenone within a concentration range of 50 microM (detection limit) to 250 microM after 12 and 24 h. Production of the Apc-mCherry fusion protein in the reporter strain is highly specific and responds to acetophenone and both enantiomers of 1-phenylethanol |
| 6.5.1.1 | DNA ligase (ATP) |
analysis |
essential reagent in studies on nucleic acid structure and metabolism. In combination with polynucleotide kinase end-group labeling, DNA ligase can be used to identify 3'- and 5'-end groups at single-strand interruptions by nearest neighbor analysis. DNA |
| 6.5.1.1 | DNA ligase (ATP) |
analysis |
a procedure for fast, sensitive and quantitative measurement of DNA ligase activity in crude cell extract |
| 6.5.1.1 | DNA ligase (ATP) |
analysis |
DNA automaton based on FokI without ligase has similar efficiency as with ligase in the context of automaton reactions. Other enzymes (HgaI, BsmFI, BbsI, and BseMII) show more discrepancy between with and without ligase |
