Journal article
Identification of Carboxylesterase, Butyrylcholinesterase, Acetylcholinesterase, Paraoxonase, and Albumin Pseudoesterase in Guinea Pig Plasma through Nondenaturing Gel Electrophoresis
Comparative medicine, Vol.68(5), pp.367-374
10/01/2018
DOI: 10.30802/AALAS-CM-18-000047
PMID: 30278860
Abstract
Drugs to protect against nerve agent toxicity are tested in animals. The current preferred small animal model is guinea pigs because their plasma bioscavenging capacity resembles that of NHP. We stained nondenaturing polyacrylamide slab gels with a variety of substrates, inhibitors, and antibodies to identify the esterases in heparinized guinea pig plasma. An intense band of carboxylesterase activity migrated behind albumin. Minor carboxylesterase bands were revealed after background activity from paraoxonase was inhibited by using EDTA. The major butyrylcholinesterase band was a disulfide-linked dimer. Incubation with the antihuman butyrylcholinesterase antibody B2 18-5 shifted the butyrylcholinesterase dimer band to slower migrating complexes. Carboxylesterases were distinguished from butyrylcholinesterase by their sensitivity to inhibition by bis-p-nitrophenyl phosphate. Acetylcholinesterase tetramers formed a complex with the antihuman acetylcholinesterase antibody HR2. Organophosphorus toxicants including cresyl saligenin phosphate, dichlorvos, and chlorpyrifos oxon irreversibly inhibited the serine esterases but not paraoxonase. Albumin pseudoesterase activity was seen in gels stained with α- or β-naphthyl acetate and fast blue RR. We conclude that guinea pig plasma has 2 types of carboxylesterase, butyrylcholinesterase dimers and 5 minor butyrylcholinesterase forms, a small amount of acetylcholinesterase tetramers, paraoxonase, and albumin pseudoesterase activity. A knockout mouse with no carboxylesterase activity in plasma is available and may prove to be a better model for studies of nerve agent toxicology than guinea pigs.
Details
- Title: Subtitle
- Identification of Carboxylesterase, Butyrylcholinesterase, Acetylcholinesterase, Paraoxonase, and Albumin Pseudoesterase in Guinea Pig Plasma through Nondenaturing Gel Electrophoresis
- Creators
- Geoffroy Napon - University of Nebraska Medical CenterAlicia J Dafferner - University of Nebraska Medical CenterAshima Saxena - Walter Reed Army Institute of ResearchOksana Lockridge - University of Nebraska Medical Center
- Resource Type
- Journal article
- Publication Details
- Comparative medicine, Vol.68(5), pp.367-374
- DOI
- 10.30802/AALAS-CM-18-000047
- PMID
- 30278860
- NLM abbreviation
- Comp Med
- ISSN
- 1532-0820
- eISSN
- 2769-819X
- Publisher
- American Association for Laboratory Animal Science
- Number of pages
- 8
- Language
- English
- Date published
- 10/01/2018
- Academic Unit
- Internal Medicine
- Record Identifier
- 9985179800102771
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