Identification of carboxylesterase, butyrylcholinesterase, acetylcholinesterase, paraoxonase, and albumin pseudoesterase in Guinea pig plasma through nondenaturing gel electrophoresis

Geoffroy Napon, Alicia J. Dafferner, Ashima Saxena, Oksana Lockridge

Research output: Contribution to journalArticle

1 Citation (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)367-374
Number of pages8
JournalComparative Medicine
Volume68
Issue number5
DOIs
StatePublished - Oct 2018

Fingerprint

Butyrylcholinesterase
Aryldialkylphosphatase
carboxylesterase
Carboxylesterase
cholinesterase
Acetylcholinesterase
acetylcholinesterase
Electrophoresis
guinea pigs
gel electrophoresis
albumins
Albumins
Guinea Pigs
Gels
Plasmas
Dimers
esterases
antibodies
Antibodies
Animals

Keywords

  • BNPP
  • Bis-nitrophenyl phosphate
  • CBDP
  • Cresyl saligenin phosphate
  • HuBChE
  • Human butyrylcholinesterase
  • Tetraisopropyl pyrophosphoramide
  • isoOMPA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • veterinary(all)

Cite this

Identification of carboxylesterase, butyrylcholinesterase, acetylcholinesterase, paraoxonase, and albumin pseudoesterase in Guinea pig plasma through nondenaturing gel electrophoresis. / Napon, Geoffroy; Dafferner, Alicia J.; Saxena, Ashima; Lockridge, Oksana.

In: Comparative Medicine, Vol. 68, No. 5, 10.2018, p. 367-374.

Research output: Contribution to journalArticle

@article{0c8cba0cde3a4895a1d936388e98266e,
title = "Identification of carboxylesterase, butyrylcholinesterase, acetylcholinesterase, paraoxonase, and albumin pseudoesterase in Guinea pig plasma through nondenaturing gel electrophoresis",
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.",
keywords = "BNPP, Bis-nitrophenyl phosphate, CBDP, Cresyl saligenin phosphate, HuBChE, Human butyrylcholinesterase, Tetraisopropyl pyrophosphoramide, isoOMPA",
author = "Geoffroy Napon and Dafferner, {Alicia J.} and Ashima Saxena and Oksana Lockridge",
year = "2018",
month = "10",
doi = "10.30802/AALAS-CM-18-000047",
language = "English (US)",
volume = "68",
pages = "367--374",
journal = "Comparative Medicine",
issn = "1532-0820",
publisher = "American Association for Laboratory Animal Science",
number = "5",

}

TY - JOUR

T1 - Identification of carboxylesterase, butyrylcholinesterase, acetylcholinesterase, paraoxonase, and albumin pseudoesterase in Guinea pig plasma through nondenaturing gel electrophoresis

AU - Napon, Geoffroy

AU - Dafferner, Alicia J.

AU - Saxena, Ashima

AU - Lockridge, Oksana

PY - 2018/10

Y1 - 2018/10

N2 - 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.

AB - 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.

KW - BNPP

KW - Bis-nitrophenyl phosphate

KW - CBDP

KW - Cresyl saligenin phosphate

KW - HuBChE

KW - Human butyrylcholinesterase

KW - Tetraisopropyl pyrophosphoramide

KW - isoOMPA

UR - http://www.scopus.com/inward/record.url?scp=85055629677&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055629677&partnerID=8YFLogxK

U2 - 10.30802/AALAS-CM-18-000047

DO - 10.30802/AALAS-CM-18-000047

M3 - Article

C2 - 30278860

AN - SCOPUS:85055629677

VL - 68

SP - 367

EP - 374

JO - Comparative Medicine

JF - Comparative Medicine

SN - 1532-0820

IS - 5

ER -