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C E Furlong

Publications and source records attributed to C E Furlong.

At least 55 records · Page 3Linked to original sources
PubMed1988

Role of genetic polymorphism of human plasma paraoxonase/arylesterase in hydrolysis of the insecticide metabolites chlorpyrifos oxon and paraoxon.

Plasma paraoxonase is a polymorphic enzyme that hydrolyzes paraoxon, the neurotoxic, active metabolite of the insecticide parathion. This enzyme is specified by at least two alleles with frequencies of about .7 and .3 among Caucasoid populations. A specific assay was developed that measured the activity of human plasma paraoxonase without interference from serum albumin which contributes significantly to the hydrolytic breakdown of paraoxon at the high pH values used in many previous assays. There was an 11-fold variation in paraoxonase activities, and the population distribution was at least bimodal. However, this specific assay did not improve the discrimination between the three genetic classes: (1) homozygotes for the low-activity allele, (2) heterozygotes, and (3) homozygotes for the high-activity allele. Chlorpyrifos oxon--the neurotoxic metabolite of the organophosphorus insecticide chlorpyrifos (Dursban)--was hydrolyzed by the same plasma fraction that hydrolyzed paraoxon. There was only four- to fivefold variability in enzyme activity, and the population distribution was unimodal. Homozygotes for low paraoxonase activity ranged over almost the entire spectrum of chlorpyrifos oxonase activity. Possible differences in susceptibility to chlorpyrifos toxicity therefore are unlikely to be predicted by the paraoxonase genotype alone. The ratio of paraoxonase over that of chlorpyrifos oxonase provided an excellent method for genetic typing of the paraoxonase polymorphism, as did the substitution of phenylacetate for chlorpyrifos as the substrate.

Alleles↗
PubMed1987

Expression of the Escherichia coli xylose isomerase gene in Saccharomyces cerevisiae.

Transformation of Saccharomyces cerevisiae by yeast expression plasmids bearing the Escherichia coli xylose isomerase gene leads to production of the protein. Western blotting (immunoblotting) experiments show that immunoreactive protein chains which comigrate with the E. coli enzyme are made in the transformant strains and that the amount produced parallels the copy number of the plasmid. When comparable amounts of immunologically cross-reactive xylose isomerase protein made in E. coli or S. cerevisiae were assayed for enzymatic activity, however, the yeast protein was at least 10(3)-fold less active.

Aldose-Ketose Isomerases↗
PubMed1984

Xylose isomerase from Escherichia coli. Characterization of the protein and the structural gene.

The gene that codes for xylose isomerase in Escherichia coli has been cloned by complementation of a xylose isomerase-negative E. coli mutant. The structural gene is 1320 nucleotides in length and codes for a protein of 440 amino acids. An additional 209 nucleotides 5' and 82 nucleotides 3' to the structural gene were also sequenced. To verify that the cloned gene encodes E. coli xylose isomerase, the enzyme was purified to homogeneity and the sequence of the first 25 amino acid residues was determined by a semimicromanual Edman procedure. These results establish that the NH2-terminal methionine of xylose isomerase is specified by an ATG which is 7 nucleotides downstream from a Shine-Dalgarno sequence.

Aldose-Ketose Isomerases↗
PubMed1984

Paraoxon hydrolysis in human serum mediated by a genetically variable arylesterase and albumin.

Gel filtration chromatography resolves human serum paraoxonase into two fractions: (1) a high molecular weight fraction that is completely inhibited by EDTA and coelutes with arylesterase (E.C.3.1.1.2); and (2) a second fraction that is closely associated with albumin, is only partially inhibited by EDTA, and has relatively little arylesterase activity under the assay conditions used. The activity of the high molecular weight fraction is stimulated by NaCl, whereas the albumin associated activity is partially inhibited by NaCl and is not present in serum derived from an analbuminemic individual. Our data suggest that albumin itself, rather than a protein bound to or cofractionating with albumin, mediates paraoxonase activity. The variation in levels of the activity of the nonalbumin, high molecular weight enzyme is responsible for the observed polymorphism of paraoxonase activity in human serum or plasma. An optimal assay of polymorphic paraoxonase activity should be based on activity measurements of the nonalbumin fraction. It is considered likely that only the nonalbumin fraction is responsible for in vivo hydrolysis of paraoxon.

Aryldialkylphosphatase↗
PubMed1983

The amino acid sequence of D-ribose-binding protein from Escherichia coli K12.

The amino acid sequence of the D-ribose-binding protein from Escherichia coli K12 was determined from the DNA sequence of the gene. Protein sequence analyses covering 80% of the protein were consistent with the sequence deduced from the DNA. The mature binding protein has 271 amino acid residues and shows substantial homology to D-galactose-binding protein. A signal peptide sequence of 23 or 25 residues was also deduced from the DNA sequence. It shows the characteristic features of prokaryotic signal peptides.

Amino Acid Sequence↗
PubMed1983

Biosynthesis of bacterial glycogen. Primary structure of Escherichia coli ADP-glucose synthetase as deduced from the nucleotide sequence of the glg C gene.

The nucleotide sequence of the glg C gene of Escherichia coli K12, coding for ADP-glucose synthetase, has been determined. The structural gene consists of 1293 base pairs, which specify a protein of 431 amino acids. The amino acid sequence deduced from the DNA sequence is consistent with the known NH2-terminal amino acid sequence and the amino acid composition of ADP-glucose synthetase. The translation start of the structural gene of glycogen synthase, glg A, starts immediately after termination of the glg C gene.

Amino Acid Sequence↗
PubMed1983

Sequence of a Drosophila DNA segment that functions in Saccharomyces cerevisiae and its regulation by a yeast promoter.

We have determined the complete DNA sequence of a segment derived from Drosophila melanogaster that complements a yeast adenine-8 (ade8) mutation. This sequence, in conjunction with transcriptional analysis, predicts that a 206 amino acid sequence accounts for complementation. We have ligated a 1.5 kb promoter segment of the yeast alcohol dehydrogenase I (ADHI) gene just upstream of the open reading frame. This construction results in ADE8+ function that responds to glucose and ethanol in a manner similar to that for the chromosomal ADHI gene, as determined by a simple quantitative color assay for ADE8+ function.

Adenine↗
PubMed1983

Plasma paraoxonase polymorphism: a new enzyme assay, population, family, biochemical, and linkage studies.

Plasma paraoxonase hydrolyzes paraoxon, the principal metabolite of the insecticide parathione. A genetic polymorphism for enzyme activity has been previously demonstrated. We describe a new assay based on the differential inhibition by EDTA of plasma paraoxonase from persons with the high-activity allele (PX*H) that suggests a trimodality of activity levels in population studies. The gene frequency of the low activity allele (PX*L) in 531 Seattle blood donors of European origin was .7207. Family studies were consistent with codominant autosomal inheritance of two alleles, PX*L (low) and PX*H (high), coding for products with different activity levels. Biochemical measurements of sera from presumed homozygotes for the two different alleles revealed minor physicochemical differences suggestive of a structural difference between the allelic products. No evidence for linkage of the paraoxonase locus with any of 19 polymorphic markers would be detected.

Adolescent↗
PubMed1982

Renaturation and identification of periplasmic proteins in two-dimensional gels of Escherichia coli.

The locations of the periplasmic proteins of Escherichia coli on standard two-dimensional gel patterns are described. The periplasmic fractions were prepared by osmotic shock of plasmolyzed whole cells and by release during EDTA-lysozyme treatment of whole cells. Within this fraction of proteins, we identify nine binding proteins (leucine-specific, glutamate-aspartate, glutamine, cystine, galactose, maltose, xylose, ribose, and arabinose) in addition to leucine-isoleucine-valine binding protein, which has been previously identified (Bloch, P. L., Phillips, T. A., and Neidhardt, F. C. (1980) J. Bacteriol. 141, 1409-1420). The identifications are based upon genetic criteria, protein induction, and comigration with purified protein. The levels of these proteins are compared in strains K12, B, and HA12 (a derivative of W). A technique was developed for renaturation of the ligand binding sites of periplasmic binding proteins in denaturing two-dimensional gels. This technique was used to demonstrate that leucine-specific and cystine binding proteins both have different isoelectric points in different strains. Renaturation was also used to demonstrate that there are two different charged forms for glutamine binding protein.

Carrier Proteins↗
PubMed1982

High resolution two-dimensional gel electrophoresis of human erythrocyte membrane proteins.

Three different two-dimensional (2-D) gel electrophoretic techniques have been modified to provide high resolution of human erythrocyte membrane proteins. The resulting gels were referenced to the established one-dimensional (1-D) sodium dodecylsulfate (SDS) gel electrophoretic profile, and the effects of endogenous proteolysis and cytosolic contamination were studied. It is concluded that in vitro proteolysis and cytosolic contamination do not contribute significantly to the patterns observed on the 2-D gels, under the conditions used for erythrocyte ghost preparation. The procedures require only small quantities of blood; as many as twenty 2-D gel profiles can be obtained from 5 ml of blood. The combination of nonequilibrium isoelectric focusing (IEF) in the first dimension, SDS electrophoresis in the second dimension, and very sensitive silver staining techniques resolves more than 250 individual protein spots. This appears to be the most useful single procedure for the analysis of red cell membrane proteins. Membrane protein profiles from patients with Duchenne muscular dystrophy, Wernicke-Korsakoff syndrome, and acanthocytosis with degeneration of the basal ganglia were compared with normal controls. The patterns for Duchenne muscular dystrophy and Wernicke-Korsakoff syndrome were not different from normal patterns. The pattern for the patient with acanthocytosis and degeneration of the basal ganglia consistently showed a high level for one protein in the 100,000 mol. wt. range.

Acantholysis↗
PubMed1981

Neutral amino acid transport during the cell cycle of cultured human lymphocytes.

Neutral amino acid (isoleucine, leucine, and valine) transport was monitored in synchronous populations of WIL-2 cells, a diploid human lymphoblastoid B cell line. Cells were synchronized by either the double thymidine block technique or by density-dependent arrest in G1. Cells synchronized by these methods showed up to a 66% decrease in amino acid transport upon progressing from the early S phase through the G2 and M phases with a concomitant increase in transport during the G1 phase. The lowest activities of amino acid transport in synchronized cells were observed during the G2 and M phases.

Amino Acids, Branched-Chain↗
PubMed1980

Reconstitution of binding protein dependent ribose transport in spheroplasts derived from a binding protein negative Escherichia coli K12 mutant and from Salmonella typhimurium.

Highly purified ribose-binding protein from Escherichia coli has been used to reconstitute binding-protein-dependent ribose transport in spheroplasts derived from a binding-protein-deficient mutant of E coli K12, and in spheroplasts derived from Salmonella typhimurium. The cross-species reconstitution was nearly as efficient as the reconstitution of the E coli strain from which the binding protein was derived. Antibody raised against the ribose binding protein completely prevented reconstitution, whereas it had no effect on whole cells. The reconstitution procedure has been improved by generating spheroplasts from cells grown in a rich medium and by reducing the background uptake in spheroplasts through a special washing procedure. Rapid purification of ribose binding protein by high pressure liquid chromatography is also described.

Bacterial Proteins↗