Human and rabbit paraoxonases: Purification, cloning, sequencing, mapping and role of polymorphism in organophosphate detoxification

C. E. Furlong, L. G. Costa, C. Hassett, R. J. Richter, J. A. Sundstrom, D. A. Adler, C. M. Disteche, C. J. Omiecinski, C. Chapline, J. W. Crabb, R. Humbert

Research output: Contribution to journalArticle

115 Scopus citations

Abstract

Human and rabbit paraoxonases/arylesterases were purified to homogeneity by chromatographic and gel electrophoretic/isofocusing procedures coupled with activity stains. N-terminal and peptide sequence analysis suggested retention of the secretion signal sequence and allowed design of oligonucleotide probes. The probes were used to isolate a 1294-bp rabbit paraoxonase cDNA clone, which, in turn, was used to isolate three human cDNA clones. Comparison of rabbit and human protein and cDNA sequences indicated a high degree of sequence conservation (∼ 85% identity) and verified that paraoxonase retains its signal sequence (except for the N-terminal Met). The rabbit cDNA encodes a protein of 359 amino acids and the human a protein of 355 amino acids. In situ hybridization demonstrated, as expected, that the paraoxonase gene maps to the long arm of human chromosome 7. Arginine at position 192 specifies high activity paraoxonase and glutamine low activity human paraoxonase. Variation in protein levels explains the variation of enzyme activity observed within a genetic class. Toxicity studies showed that raising rat plasma paraoxonase levels by i.v. administration of partially purified rabbit paraoxonase protected animals against cholinesterase inhibition by paraoxon and chlorpyrifos oxon. Protection correlated with the relative rates of hydrolysis of these two compounds.

Original languageEnglish (US)
Pages (from-to)35-48
Number of pages14
JournalChemico-Biological Interactions
Volume87
Issue number1-3
DOIs
StatePublished - Jun 1993

All Science Journal Classification (ASJC) codes

  • Toxicology

Fingerprint Dive into the research topics of 'Human and rabbit paraoxonases: Purification, cloning, sequencing, mapping and role of polymorphism in organophosphate detoxification'. Together they form a unique fingerprint.

  • Cite this

    Furlong, C. E., Costa, L. G., Hassett, C., Richter, R. J., Sundstrom, J. A., Adler, D. A., Disteche, C. M., Omiecinski, C. J., Chapline, C., Crabb, J. W., & Humbert, R. (1993). Human and rabbit paraoxonases: Purification, cloning, sequencing, mapping and role of polymorphism in organophosphate detoxification. Chemico-Biological Interactions, 87(1-3), 35-48. https://doi.org/10.1016/0009-2797(93)90023-R