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The Journal of Experimental Zoology Apr 1967
Topics: Animals; Electrophoresis; Histocytochemistry; Isoenzymes; L-Lactate Dehydrogenase; Male; Mice; Organ Size; Spectrophotometry; Spermatozoa; Testis
PubMed: 6068185
DOI: 10.1002/jez.1401640302 -
Clinical Chemistry Sep 1988
Topics: Cold Temperature; Drug Stability; Humans; Isoenzymes; L-Lactate Dehydrogenase; Lung Neoplasms
PubMed: 3416478
DOI: No ID Found -
Journal of Biochemistry Jan 1991L-Lactate dehydrogenase of Thermus caldophilus GK24 was purified from Escherichia coli containing an overexpression plasmid. The enzyme was crystallized from...
L-Lactate dehydrogenase of Thermus caldophilus GK24 was purified from Escherichia coli containing an overexpression plasmid. The enzyme was crystallized from polyethylene glycol 6000 solutions without ligands by the hanging drop vapor diffusion method. Two forms of crystals were obtained. The crystals grown at pH 6.0 were characterized by means of an X-ray diffraction experiment, while those grown at pH 6.5 and 7.0 did not give detectable diffraction spots. The crystals grown at pH 6.0 belonged to monoclinic space group P2(1), the cell dimensions being a = 54.8 A, b = 138.2A, c = 86.1 A, and beta = 93.3 degrees. These crystals diffract to beyond 2.5 A spacing and are stable on X-ray irradiation.
Topics: Allosteric Regulation; Crystallization; L-Lactate Dehydrogenase; Molecular Structure; Thermus; X-Ray Diffraction
PubMed: 2016275
DOI: No ID Found -
Isozymes 1983
Topics: Cells, Cultured; Erythrocytes; Fibroblasts; Glycolysis; Humans; Ischemia; Isoenzymes; L-Lactate Dehydrogenase; Leukocytes; Macromolecular Substances; Muscles; Physical Exertion; Skin
PubMed: 6642993
DOI: No ID Found -
Nature Apr 1968
Topics: Animals; Biological Evolution; Bone Marrow; Bone Marrow Cells; Electrophoresis; L-Lactate Dehydrogenase; Molecular Biology; Rodentia; Starch
PubMed: 5649670
DOI: 10.1038/218341a0 -
Biotechnology Progress 2009We used zeolite beta as an adsorbing matrix to refold recombinant lactate dehydrogenase (LDH) protein collected as an insoluble aggregate from a bacterial expression...
We used zeolite beta as an adsorbing matrix to refold recombinant lactate dehydrogenase (LDH) protein collected as an insoluble aggregate from a bacterial expression system. The adsorption isotherm revealed that 1 g of zeolite adsorbed 200 mg of denatured LDH solubilized with a buffer containing 6 M of guanidine hydrochloride. The pH of the buffer had little effect on the adsorption, but this property was abolished by preincubation of the zeolite with polyethylene glycol (PEG) in a weight ratio of 1:10. These data suggest that the adsorption of LDH depends on the hydrophobicity of the zeolite surface, and that the adsorption of PEG to zeolite is sufficient to release LDH from its surface. LDH was thus released by refolding buffer containing PEG and arginine, and soluble LDH was obtained in its active enzymatic form. The addition of arginine dramatically increased the yield of LDH in a dose-dependent manner. The overall refolding efficiency was optimized to 35%.
Topics: L-Lactate Dehydrogenase; Protein Folding; Recombinant Proteins; Zeolites
PubMed: 19224588
DOI: 10.1002/btpr.107 -
FEMS Microbiology Letters Mar 1991The NAD-dependent D-lactate dehydrogenase from Lactobacillus bulgaricus has been purified to homogeneity. This enzyme was a dimer made of two identical chains of...
The NAD-dependent D-lactate dehydrogenase from Lactobacillus bulgaricus has been purified to homogeneity. This enzyme was a dimer made of two identical chains of molecular mass 37,000. Saturation by either substrate was hyperbolic, with Km values of 50 microM for NADH and 1 mM for pyruvate. The specific activity was 2200 units/mg and was not affected by the presence of fructose-1,6-bisphosphate, Mn2+ ions, ATP or ADP. The amino-terminal sequence determined on 50 residues showed no significant homology with known lactate dehydrogenases, suggesting that the D-lactate dehydrogenase from L. bulgaricus could not be evolutionarily related to the family of NAD-dependent L-lactate dehydrogenases.
Topics: Amino Acid Sequence; Biological Evolution; Electrophoresis, Polyacrylamide Gel; L-Lactate Dehydrogenase; Lactobacillus; Molecular Sequence Data; NAD; Pyruvates
PubMed: 2044942
DOI: 10.1016/0378-1097(91)90533-g -
Molecular Biology and Evolution Sep 2022We unveil the intimate relationship between protein dynamics and allostery by following the trajectories of model proteins in their conformational and sequence spaces....
Protein Conformational Space at the Edge of Allostery: Turning a Nonallosteric Malate Dehydrogenase into an "Allosterized" Enzyme Using Evolution-Guided Punctual Mutations.
We unveil the intimate relationship between protein dynamics and allostery by following the trajectories of model proteins in their conformational and sequence spaces. Starting from a nonallosteric hyperthermophilic malate dehydrogenase, we have tracked the role of protein dynamics in the evolution of the allosteric capacity. Based on a large phylogenetic analysis of the malate (MalDH) and lactate dehydrogenase (LDH) superfamily, we identified two amino acid positions that could have had a major role for the emergence of allostery in LDHs, which we targeted for investigation by site-directed mutagenesis. Wild-type MalDH and the single and double mutants were tested with respect to their substrate recognition profiles. The double mutant displayed a sigmoid-shaped profile typical of homotropic activation in LDH. By using molecular dynamics simulations, we showed that the mutations induce a drastic change in the protein sampling of its conformational landscape, making transiently T-like (inactive) conformers, typical of allosteric LDHs, accessible. Our data fit well with the seminal key concept linking protein dynamics and evolvability. We showed that the selection of a new phenotype can be achieved by a few key dynamics-enhancing mutations causing the enrichment of low-populated conformational substates.
Topics: Allosteric Regulation; Amino Acids; L-Lactate Dehydrogenase; Malate Dehydrogenase; Malates; Mutation; Phylogeny
PubMed: 36056899
DOI: 10.1093/molbev/msac186 -
Journal of Pharmaceutical Sciences Nov 2007The effects of eight cyclodextrins (CDs) on the stability of freeze-dried lactate dehydrogenase (LDH) were investigated. Five low-molecular weight saccharides were used...
The effects of eight cyclodextrins (CDs) on the stability of freeze-dried lactate dehydrogenase (LDH) were investigated. Five low-molecular weight saccharides were used as a reference. In the samples without saccharides, LDH activity was decreased by freeze-drying. The LDH activities of the samples with 2-hydroxypropylated CDs (HP-CDs) such as 2-hydroxypropyl-beta-cyclodextrin (HP-beta-CD) showed only a small decrease and were more effective protein stabilizers than low-molecular weight saccharides. HP-beta-CD more effectively stabilized LDH than trehalose, at a sugar concentration of less than 1.0 w/w% and furthermore, a close relationship existed between the highly stabilizing effect and the degree of substitution of polar substituents.
Topics: Cyclodextrins; Enzyme Stability; Freeze Drying; L-Lactate Dehydrogenase; Molecular Weight; Polysaccharides
PubMed: 17828735
DOI: 10.1002/jps.20847 -
Biological Trace Element Research Aug 2009Lactate dehydrogenase (LDH, EC1.1.1.27), widely expressed in the heart, liver, and other tissues, plays an important role in glycolysis and glyconeogenesis. The activity...
Lactate dehydrogenase (LDH, EC1.1.1.27), widely expressed in the heart, liver, and other tissues, plays an important role in glycolysis and glyconeogenesis. The activity of LDH is often altered upon inflammatory responses in animals. Nano-TiO(2) was shown to provoke various inflammatory responses both in rats and mice; however, the molecular mechanism by which TiO(2) exerts its toxicity has not been completely understood. In this report, we investigated the mechanisms of nano-anatase TiO(2) (5 nm) on LDH activity in vitro. Our results showed that LDH activity was greatly increased by low concentration of nano-anatase TiO(2), while it was decreased by high concentration of nano-anatase TiO(2). The spectroscopic assays revealed that the nano-anatase TiO(2) particles were directly bound to LDH with mole ratio of [nano-anatase TiO(2)] to [LDH] was 0.12, indicating that each Ti atom was coordinated with five oxygen/nitrogen atoms and a sulfur atoms of amino acid residues with the Ti-O(N) and Ti-S bond lengths of 1.79 and 2.41 A. We postulated that the bound nano-anatase TiO(2) altered the secondary structure of LDH, created a new metal ion-active site for LDH, and thereby enhanced LDH activity.
Topics: Animals; Coloring Agents; Enzyme Activation; Heart; L-Lactate Dehydrogenase; Myocardium; Nanoparticles; Rats; Titanium
PubMed: 19214398
DOI: 10.1007/s12011-009-8326-9