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Parasitology Research Jun 2006From a Clonorchis sinensis adult worm cDNA library, we isolated a cDNA clone encoding a novel lactate dehydrogenase (LDH) gene which encoded a putative protein with a...
From a Clonorchis sinensis adult worm cDNA library, we isolated a cDNA clone encoding a novel lactate dehydrogenase (LDH) gene which encoded a putative protein with a predicted molecular weight of 35.6 kDa. The optimum pH and temperature for the enzyme were 7.5 and 50 degrees C in the pyruvate reduction while 11 and 80 degrees C in the lactate oxidation reaction, respectively. CsLDH showed no substrate inhibition by high lactate and NAD(+) concentration, and the optimal pyruvate and optimal NADH concentrations were 10 and 0.5 mmol/l, respectively. The relative activities of these 2-oxocarboxylic acids were pyruvic acid>2-ketobutyrate>oxalacetic acid>alpha-ketoglutaric acid>phenylpyruvate. The cofactor 3-acetylpyridine adenine dinucleotide was much more effective than NAD(+). The cofactor analogs in which the nicotinamide ring is replaced by 3-pyridinealdehyde were lower activity cofactors, while the nicotinamide ring is replaced by nicotinic acid or thionicotinamide which is not a cofactor to CsLDH. The succinic acid and malic acid are not substrates of CsLDH. Cu(2+), Fe(2+), and Zn(2+) greatly inhibited the CsLDH activity both in the direction of pyruvate reduction and in the direction of lactate oxidation. The inhibition of CsLDH by gossypol may make gossypol a potential therapy drug or a lead compound for C. sinensis. Accordingly, the CsLDH may be a novel potential drug target.
Topics: Amino Acid Sequence; Animals; Base Sequence; Carboxylic Acids; Clonorchis sinensis; Enzyme Inhibitors; Gossypol; Helminth Proteins; Hydrogen-Ion Concentration; L-Lactate Dehydrogenase; Molecular Sequence Data; Molecular Weight; Oxidation-Reduction; Sequence Alignment; Substrate Specificity; Temperature
PubMed: 16479375
DOI: 10.1007/s00436-005-0125-4 -
The Journal of Biological Chemistry Aug 1974
Comparative Study
Topics: Animals; Binding Sites; Carbon Radioisotopes; Cellulose; Chromatography, Gel; Chromatography, Ion Exchange; Cycloheximide; Dactinomycin; Electrophoresis; Freezing; Half-Life; Isoenzymes; Kinetics; L-Lactate Dehydrogenase; Leucine; Liver; Mathematics; Myocardium; Protein Binding; Rats; Spectrophotometry, Ultraviolet; Time Factors; Tritium
PubMed: 4846754
DOI: No ID Found -
Zeitschrift Fur Klinische Chemie Und... May 1968
Topics: Electrophoresis; Glycolysis; Humans; Isoenzymes; Kidney Tubules; L-Lactate Dehydrogenase; Methods
PubMed: 5709550
DOI: 10.1515/cclm.1968.6.3.132 -
Biochemistry Jun 1986Solutions of porcine lactate dehydrogenase of micromolar concentration kept at 4 degrees C for several days lose the greater part of their enzymic activity but recover...
Solutions of porcine lactate dehydrogenase of micromolar concentration kept at 4 degrees C for several days lose the greater part of their enzymic activity but recover it when returned to room temperature. The rate of spoiling decreases and the rate of recovery increases with the concentration of the solutions. The decrease in tetramer stability in the cold is shown by experiments of pressure dissociation at various temperatures and confirmed because isozyme hybridization occurs in parallel with the inactivation at low temperature but is absent at room temperature. Cold-inactivated solutions contain tetramers that dissociate much more readily than those of the fully active solutions. It is postulated that cryoinactivation, like pressure inactivation, takes place through a cycle of dissociation, conformational drift [King, L., & Weber, G. (1986) Biochemistry (second paper of three in this issue)] and reassociation into inactive tetramers.
Topics: Animals; Freezing; Isoenzymes; Kinetics; L-Lactate Dehydrogenase; Myocardium; Protein Conformation; Swine
PubMed: 3718950
DOI: 10.1021/bi00360a024 -
Genetica May 1987A new electrophoretic variant of the lactate dehydrogenase B subunit was found in the erythrocytes of the COP strain of the rat. The location of the band after the...
A new electrophoretic variant of the lactate dehydrogenase B subunit was found in the erythrocytes of the COP strain of the rat. The location of the band after the electrophoresis suggested a product of the structural gene for the B subunit. Two alleles that regulated the high amount (Ldh-2a) or the low amount (Ldh-2b) of the B subunit were found and segregated in Mendelian fashion. The activity was regulated by the closely linked (less than 1 cM) regulatory gene Ldr-1.
Topics: Alleles; Animals; Electrophoresis, Polyacrylamide Gel; Genes; Genes, Regulator; Genetic Linkage; Genetic Variation; Isoenzymes; L-Lactate Dehydrogenase; Polymorphism, Genetic; Protein Conformation; Rats
PubMed: 3505879
DOI: 10.1007/BF00126979 -
Biofizika 1999A new method for encapsulating enzymes by multilayer polyelectrolyte coating is proposed. The method consists in a stepwise adsorption of polyelectrolytes from solution...
A new method for encapsulating enzymes by multilayer polyelectrolyte coating is proposed. The method consists in a stepwise adsorption of polyelectrolytes from solution onto protein aggregates formed by salting out the proteins in highly concentrated salt solutions. Polystyrene sulfonate and fluorescence-labeled polyalylamine were used for capsule formation. The size of lactate dehydrogenase aggregates covered by four layer pairs of electrolytes was 1-5 microns, as indicated by fluorescence microscopy. The catalytic characteristics and stability of pig muscle lactate dehydrogenase (EC 1.1.1.13) incapsulated in multilayer electrolyte complex obtained by this method were studied. It was found that the affinity of the substrate pyruvate for the enzyme in the polyelectrolyte complex (K(M)) did not essentially change as compared with the free enzyme. Incapsulated lactate dehydrogenase showed the following features that distinguish it from the free form: (1) the lifetime in diluted solutions increases from 30 min (without capsules) to 1-2 days (in capsules); (2) a higher stability to basic denaturation (up to pH 10); and (3) the absence of substrate inhibition of enzyme in the polyelectrolyte complex. The changes in the catalytic characteristics of incapsulated lactate dehydrogenase are discussed in terms of an increase in effective pK values of amino acid perturbed by polyelectrolyte coating of enzyme.
Topics: Animals; Catalysis; Electrolytes; Hydrogen-Ion Concentration; Isoenzymes; L-Lactate Dehydrogenase; Muscles; Protein Denaturation; Substrate Specificity; Swine
PubMed: 10624520
DOI: No ID Found -
Memorias Do Instituto Oswaldo Cruz 1998
Topics: Animals; L-Lactate Dehydrogenase; Life Cycle Stages; Male; Schistosoma mansoni
PubMed: 9921351
DOI: 10.1590/s0074-02761998000700035 -
Comparative Biochemistry and... 19831. Previous work showed that seven out of 13 Xenopus species and subspecies possess a simple pattern of 5 LDH isozymes as it is common to most vertebrates. 2. The...
1. Previous work showed that seven out of 13 Xenopus species and subspecies possess a simple pattern of 5 LDH isozymes as it is common to most vertebrates. 2. The remaining six species with more complicated patterns are further analyzed in the present paper. 3. X. l. laevis, X. vestitus and X. wittei are characterized by the appearance of an anodal "sixth band". Two explanations are given. 4. A comparison of organs in X. borealis reveals the bands additional to the five banded pattern as secondary isozymes. 5. The five bands in the A4 region in X. ruwenzoriensis, combinations of A subunits in nature as demonstrated by treatment with antibodies against the purified A4 homopolymer, must be considered as being the result of a duplicated A. locus. 6. The same is possibly true for the closely spaced cathodal bands in X. fraseri. 7. The occurrence of only few expressed duplicated loci among the genus Xenpus indicates progressed diploidization within this genus.
Topics: Animals; Isoenzymes; L-Lactate Dehydrogenase; Species Specificity; Xenopus
PubMed: 6861472
DOI: 10.1016/0305-0491(83)90134-7 -
Analytical Sciences : the International... 2010Kinetic analyses of lactate-dehydrogenase (LD)-coupled alanine transaminase (ALT) reaction processes were investigated for measuring ALT by an integration strategy. For...
Kinetic analyses of lactate-dehydrogenase (LD)-coupled alanine transaminase (ALT) reaction processes were investigated for measuring ALT by an integration strategy. For measuring ALT by a kinetic analysis of an LD-coupled ALT reaction curve, candidate reaction curves were calculated via iterative numerical integration of the differential velocity equations to execute a weighted nonlinear-least-square-fitting. To realize the integration strategy, the conventional initial-velocity method was used if the ALT activities were below 25 U/L; otherwise, kinetic analyses of the reaction curves were employed. Of the reaction curves recorded at 10-s intervals, kinetic analyses gave ALT activities resistant to deviations in the LD kinetic parameters. The integration strategy yielded a higher value of the lower limit, but an upper limit of over 100 U/L by simulations and over 75 U/L with purified ALT. Also, its intra-run relative standard deviations were below 9% for 0.50 U/L ALT and below 5% for final 1 to 65 U/L ALT. The integration strategy gave consistent ALT activities in clinical sera. Hence, this new approach for kinetic analyses of ALT reaction processes and the integration strategy were effective to measure ALT.
Topics: Alanine Transaminase; Kinetics; L-Lactate Dehydrogenase
PubMed: 21079351
DOI: 10.2116/analsci.26.1193 -
Journal of Immunoassay 1988Four monoclonal antibodies (Mab) derived from mice immunized with lactate dehydrogenase 5 (LDH5) react strongly with LDH5, but weakly with LDH2 which contains a single...
Four monoclonal antibodies (Mab) derived from mice immunized with lactate dehydrogenase 5 (LDH5) react strongly with LDH5, but weakly with LDH2 which contains a single subunit of type M. Experimental evidence suggest that these antibodies are directed to an antigenic determinant in the interface between two subunits of type M. A sandwich ELISA procedure was devised, using these Mabs to identify and quantify LDH5. The procedure involves immobilization of one of these Mabs by its adsorption onto polyclonal anti-mouse IgG coated polystyrene plates, adsorption of LDH5, its identification by the same Mab as that used in the immobilization step, and finally color development by an enzyme labeled rabbit anti-mouse IgG antiserum. The method enables LDH5 to be assayed at a concentration range of 0-5 micrograms/ml.
Topics: Antibodies, Monoclonal; Antibody Specificity; Enzyme-Linked Immunosorbent Assay; Humans; Isoenzymes; L-Lactate Dehydrogenase
PubMed: 3360921
DOI: 10.1080/01971528808053209