Origin and dispersal of atypical aldehyde dehydrogenase ALDH2*487Lys

The East Asian respond with a marked facial flushing and mild to moderate symptoms of intoxication after drinking the amounts of alcohol that has no detectable effect on European. The alcohol sensitivity in Orientals is due to a delayed oxidation of aceta

Comparative electrophoretic patterns of lactase dehydrogenase and malate dehydrogenase in five Lake Victoria cichlid species

Biochemical techniques designed to compare species on the basis of protein differences were started by NUTTALL (1904) who used immunological methods to compare the serum of humans with that of other primates. Since then more refined techniques have led to better results at the protein level in taxonomy, The analyses of proteins are considered to be the simplest indirect approach to understanding the structure and function of the genetic material, deoxyribonucleic acid (DNA). Interest in these analyses arises because of the close relationship between protein structure and gene structure. Thus by comparing the properties of homologous proteins from different taxa one is in essence comparins their genes (GORMAN er al., 1971). It is now an established fact that genetic information coded in molecules of DNA is translated through a series of reactions in the structure of proteins which form the principal morphological units of the animal body at the molecular level of organization (SIBLEY, 1952). A convenient method of comparing molecular differences between species is to measure the electrophoretic mobility of proteins in a starch gel medium (ASPINWALL and TSUYUKI, 1968) or acrylamide gel (RAYMOND and WEINTRAUB, 1959; BOUCK and BALL, 1968). Proteins with enzymatic properties can be compared on the basis of catalytic activity in the presence or absence of inhibitors (KAPLAN et al., 1959); BAILEY et al., t 1970). A combination of gel electrophoresis and histochemical enzyme detection techniques (HUNTER and MARKERT, 1957) makes it possible to combine electrophoretic mobility anti catalytic activity comparison, Enzyme patterns exhibited in starch gel or acrylamide gel have been used to classify different species. BOUCK and BALL (1968)working with lactate dehydrogenase in species of Trout found that each Trout species had LDH pattern characterbtic of that species. ASPINIWALL and TSUYUKI (1968) used muscle protein electrophoretic patterns to identify hybrid fishes. TSUYUKI and ROBERTS (1963) and TSUYUKI et al. (1964-65) found that myogen protein patterns in fishes were species specific. The myogen patterns within one family were remarkably parallel with the existing morphometric classification and these patterns constituted a single criterion by which the fishes could be identified. The fish used in these investigations were collected from shallow waters (10 metres) of Lake Victoria in two areas, Jinja and Kisumu, using gillnets and beach-seines. The study included ten specimens of each of the following specIes: (l) Haplochromis michaeli (2) Haploehromis obems (3) Astatoreochromis ulluaudi (4) Tilapia zillii and (5) Tilapia nilotica.

Characterization of the Rana grylio virus 3 beta-hydroxysteroid dehydrogenase and its novel role in suppressing virus-induced cytopathic effect

The 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) isoenzymes play a key role in cellular steroid hormone synthesis. Here, a 3 beta-HSD gene homolog,was cloned from Rana grylio virus (RGV), a member of family Iridoviridae. RGV 3 beta-HSD gene has 1068 bp, encoding a 355 aa predicted protein. Transcription analyses showed that RGV 3 beta-HSD gene was transcribed immediate-early during infection from an initiation site 19 nucleotides upstream of the translation start site. Confocal microscopy revealed that the 3 beta-HSD-EGFP fusion protein was exclusively colocalized with the mitochondria marker (pDsRed2-Mito) in EPC cells. Upon morphological observation and MTT assay, it was revealed that overexpression of RGV 3 beta-HSD in EPC cells could apparently suppress RGV-induced cytopathic effect (CPE). The present studies indicate that the RGV immediate-early 3 beta-HSD gene encodes a mitochondria-localized protein, which has a novel role in suppressing virus-induced CPE. All these suggest that RGV 3 beta-HSD might be a protein involved in host-virus interaction. @ 2006 Elsevier Inc. All rights reserved.

Cloning and characterization of the gene for UDPGlc dehydrogenase from the cyanobacterium, Microcystis aeruginosa FACHB 905

Using degenerate primers based on conserved regions of the UDP-glucose dehydrogenase (UDPGDH) gene, an initial 476-bp DNA fragment was amplified from the water-bloom forming cyanobacterium, Microcystis aeruginosa FACHB 905. TAIL-PCR and ligation-mediated PCR were used to amplify the flanking regions to isolate an about 2.5-kb genomic DNA fragment. Sequence analysis revealed an ORF encoding a putative 462 amino acid protein, designated Mud for Microcystis UDPGDH. The Mud amino acid sequence is closely related to UDPGDH sequences from cyanobacterium Synechocystis PCC6803 (73% identity, 81% similarity), and bacterium Bacillus subtilis (51% identity and 67% similarity). The cloned mud gene was expressed in Escherichia coli using the pGEX-4T-1 fusion expression vector system to generate a GST-Mud fusion protein that exhibited UDPGDH activity. The cytosolic fraction of M aeruginosa FACHB 905 was subjected to Western analysis with an anti-Mud antibody, which revealed a single band of approximately 49 kD, consistent with the deduced molecular mass of the enzyme. The Mud protein could thus be characterized as a UDP-glucose dehydrogenase, which was a key enzyme for polysaccharide synthesis and has, for the first time, been studied in algae.

Highly ordered mesoporous carbons as electrode material for the construction of electrochemical dehydrogenase- and oxidase-based biosensors

In this work, the excel lent catalytic activity of highly ordered mesoporous carbons (OMCs) to the electrooxidation of nicotinamide adenine dinucleotide (NADH) and hydrogen peroxide (H2O2) was described for the construction of electrochemical alcohol dehydrogenase (ADH) and glucose oxidase (GOD)-based biosensors.

A novel alcohol dehydrogenase biosensor based on solid-state electrogenerated chemiluminescence by assembling dehydrogenase to Ru(bpy)(3)(2+)-Au nanoparticles aggregates

Based on electrogenerated chemiluminescence (ECL), a novel method for fabrication of alcohol dehydrogenase (ADH) biosensor by self-assembling ADH to Ru(bpy)(3)(2+) -AuNPs aggregates (Ru-AuNPs) on indium tin oxide (ITO) electrode surface has been developed. Positively charged Ru(bpy)(3)(2+) could be immobilized stably on the electrode surface with negatively charged AuNPs in the form of aggregate via electrostatic interaction. On the other hand, AuNPs are favourable candidates for the immobilization of enzymes because amine groups and cysteine residues in the enzymes are known to bind strongly with AuNPs. Moreover, AuNPs can act as tiny conduction centers to facilitate the transfer of electrons. Such biosensor combined enzymatic selectivity with the sensitivity of ECL detection for quantification of enzyme substrate, and it displayed wide linear range, high sensitivity and good stability.

Electrogenerated chemiluminescence biosensor based on Ru(bpy)(3)(2+) and dehydrogenase immobilized in sol-gel/chitosan/poly(sodium 4-styrene sulfonate) composite material

A new electrogenerated chemiluminescence biosensor was fabricated by immobilizing ECL reagent Ru(bPY)(3)(2+) and alcohol dehydrogenase in sol-gel/chitosan/poly(sodium 4-styrene sulfonate) (PSS) organically modified composite material. The component PSS was used to immobilize ECL reagent Ru(bpy)(3)(2+) by ion-exchange, while the addition of chitosan was to prevent the cracking of conventional sol-gel-derived glasses and provide biocompatible microenvironment for alcohol dehydrogenase. Such biosensor combined enzymatic selectivity with the sensitivity of ECL detection for quantification of enzyme substrate and it was much simpler than previous double-layer design. The detection limit was 9.3 x 10(-6) M for alcohol (S/N = 3) with a linear range from 2.79 x 10(-5) to 5.78 x 10(-2) M. With ECL detection, the biosensor exhibited wide linear range, high sensitivity and good stability.

Effects of rare earth ions and calmodulin on the activity of lactate dehydrogenase

In this paper, the effects of rare earth ions (La3+, Eu3+, Dy3+, Yb3+) and their complexes with calmodulin on the activity of lactate dehydrogenase (LDH) were investigated. The results reveal that whether binding with calmodulin or not, rare earth ions show a minor activation effects on LDH when their concentrations are less than 3 mu mol (.) L-1, but indicate some strong inhibitory effects on LDH activity when the concentrations are above 5 mu mol (.) L-1. Calmodulin, which is a calcium-dependent regulator, can stimulate LDH activity and release the inhibitory effects of rare earth ion. Diethylenetriamine pentaacetic acid(DTPA) and its derivatives bisdimethylamide-diethylenetriamine pentaacetic acid (DTPA-BDMA), bisisonicotinyl-diethylenetriamine pentaacetic acid (DTPA-BIN), which are often used as ligands to metal ions, inhibit LDH activity when their concentrations are above 5 mu mol (.) L-1. Calmodulin can also release their inhibitory effects at the same time.