Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry

Eric S Peeples, Lawrence M Schopfer, Ellen G. Duysen, Reggie Spaulding, Troy Voelker, Charles M. Thompson, Oksana Lockridge

Research output: Contribution to journalArticle

111 Citations (Scopus)

Abstract

The classical laboratory tests for exposure to organophosphorus toxicants (OP) are inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in blood. In a search for new biomarkers of OP exposure, we treated mice with a biotinylated organophosphorus agent, FP-biotin. The biotinylated proteins in muscle were purified by binding to avidin-Sepharose, separated by gel electrophoresis, digested with trypsin, and identified from their fragmentation patterns on a quadrupole time-of-flight mass spectrometer. Albumin and ES1 carboxylesterase (EC 3.1.1.1) were found to be major targets of FP-biotin. These FP-biotinylated proteins were also identified in mouse plasma by comparing band patterns on nondenaturing gels stained for albumin and carboxylesterase activity, with band patterns on blots hybridized with Streptavidin Alexa-680. Two additional FP-biotin targets, AChE (EC 3.1.1.7) and BChE (EC 3.1.1.8), were identified in mouse plasma by finding that enzyme activity was inhibited 50-80%. Mouse plasma contained eight additional FP-biotinylated bands whose identity has not yet been determined. In vitro experiments with human plasma showed that chlorpyrifos oxon, echothiophate, malaoxon, paraoxon, methyl paraoxon, diazoxon, diisopropylfluorophosphate, and dichlorvos competed with FP-biotin for binding to human albumin. Though experiments with purified albumin have previously shown that albumin covalently binds OP, this is the first report of OP binding to albumin in a living animal. Carboxylesterase is not a biomarker in man because humans have no carboxylesterase in blood. It is concluded that OP bound to albumin could serve as a new biomarker of OP exposure in man.

Original languageEnglish (US)
Pages (from-to)303-312
Number of pages10
JournalToxicological Sciences
Volume83
Issue number2
DOIs
StatePublished - Feb 1 2005

Fingerprint

Biomarkers
Mass spectrometry
Albumins
Mass Spectrometry
Carboxylesterase
Butyrylcholinesterase
Acetylcholinesterase
Plasmas
Blood
Gels
Dichlorvos
Paraoxon
Plasma (human)
Isoflurophate
Streptavidin
Avidin
Muscle Proteins
Cholinesterases
Enzyme activity
Mass spectrometers

Keywords

  • Acetylcholinesterase
  • Albumin
  • Butyrylcholinesterase
  • FP-biotin
  • Organophosphate

ASJC Scopus subject areas

  • Toxicology

Cite this

Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry. / Peeples, Eric S; Schopfer, Lawrence M; Duysen, Ellen G.; Spaulding, Reggie; Voelker, Troy; Thompson, Charles M.; Lockridge, Oksana.

In: Toxicological Sciences, Vol. 83, No. 2, 01.02.2005, p. 303-312.

Research output: Contribution to journalArticle

Peeples, Eric S ; Schopfer, Lawrence M ; Duysen, Ellen G. ; Spaulding, Reggie ; Voelker, Troy ; Thompson, Charles M. ; Lockridge, Oksana. / Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry. In: Toxicological Sciences. 2005 ; Vol. 83, No. 2. pp. 303-312.
@article{006d52e96823456a8d587dfa1affb8b3,
title = "Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry",
abstract = "The classical laboratory tests for exposure to organophosphorus toxicants (OP) are inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in blood. In a search for new biomarkers of OP exposure, we treated mice with a biotinylated organophosphorus agent, FP-biotin. The biotinylated proteins in muscle were purified by binding to avidin-Sepharose, separated by gel electrophoresis, digested with trypsin, and identified from their fragmentation patterns on a quadrupole time-of-flight mass spectrometer. Albumin and ES1 carboxylesterase (EC 3.1.1.1) were found to be major targets of FP-biotin. These FP-biotinylated proteins were also identified in mouse plasma by comparing band patterns on nondenaturing gels stained for albumin and carboxylesterase activity, with band patterns on blots hybridized with Streptavidin Alexa-680. Two additional FP-biotin targets, AChE (EC 3.1.1.7) and BChE (EC 3.1.1.8), were identified in mouse plasma by finding that enzyme activity was inhibited 50-80{\%}. Mouse plasma contained eight additional FP-biotinylated bands whose identity has not yet been determined. In vitro experiments with human plasma showed that chlorpyrifos oxon, echothiophate, malaoxon, paraoxon, methyl paraoxon, diazoxon, diisopropylfluorophosphate, and dichlorvos competed with FP-biotin for binding to human albumin. Though experiments with purified albumin have previously shown that albumin covalently binds OP, this is the first report of OP binding to albumin in a living animal. Carboxylesterase is not a biomarker in man because humans have no carboxylesterase in blood. It is concluded that OP bound to albumin could serve as a new biomarker of OP exposure in man.",
keywords = "Acetylcholinesterase, Albumin, Butyrylcholinesterase, FP-biotin, Organophosphate",
author = "Peeples, {Eric S} and Schopfer, {Lawrence M} and Duysen, {Ellen G.} and Reggie Spaulding and Troy Voelker and Thompson, {Charles M.} and Oksana Lockridge",
year = "2005",
month = "2",
day = "1",
doi = "10.1093/toxsci/kfi023",
language = "English (US)",
volume = "83",
pages = "303--312",
journal = "Toxicological Sciences",
issn = "1096-6080",
publisher = "Oxford University Press",
number = "2",

}

TY - JOUR

T1 - Albumin, a new biomarker of organophosphorus toxicant exposure, identified by mass spectrometry

AU - Peeples, Eric S

AU - Schopfer, Lawrence M

AU - Duysen, Ellen G.

AU - Spaulding, Reggie

AU - Voelker, Troy

AU - Thompson, Charles M.

AU - Lockridge, Oksana

PY - 2005/2/1

Y1 - 2005/2/1

N2 - The classical laboratory tests for exposure to organophosphorus toxicants (OP) are inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in blood. In a search for new biomarkers of OP exposure, we treated mice with a biotinylated organophosphorus agent, FP-biotin. The biotinylated proteins in muscle were purified by binding to avidin-Sepharose, separated by gel electrophoresis, digested with trypsin, and identified from their fragmentation patterns on a quadrupole time-of-flight mass spectrometer. Albumin and ES1 carboxylesterase (EC 3.1.1.1) were found to be major targets of FP-biotin. These FP-biotinylated proteins were also identified in mouse plasma by comparing band patterns on nondenaturing gels stained for albumin and carboxylesterase activity, with band patterns on blots hybridized with Streptavidin Alexa-680. Two additional FP-biotin targets, AChE (EC 3.1.1.7) and BChE (EC 3.1.1.8), were identified in mouse plasma by finding that enzyme activity was inhibited 50-80%. Mouse plasma contained eight additional FP-biotinylated bands whose identity has not yet been determined. In vitro experiments with human plasma showed that chlorpyrifos oxon, echothiophate, malaoxon, paraoxon, methyl paraoxon, diazoxon, diisopropylfluorophosphate, and dichlorvos competed with FP-biotin for binding to human albumin. Though experiments with purified albumin have previously shown that albumin covalently binds OP, this is the first report of OP binding to albumin in a living animal. Carboxylesterase is not a biomarker in man because humans have no carboxylesterase in blood. It is concluded that OP bound to albumin could serve as a new biomarker of OP exposure in man.

AB - The classical laboratory tests for exposure to organophosphorus toxicants (OP) are inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activity in blood. In a search for new biomarkers of OP exposure, we treated mice with a biotinylated organophosphorus agent, FP-biotin. The biotinylated proteins in muscle were purified by binding to avidin-Sepharose, separated by gel electrophoresis, digested with trypsin, and identified from their fragmentation patterns on a quadrupole time-of-flight mass spectrometer. Albumin and ES1 carboxylesterase (EC 3.1.1.1) were found to be major targets of FP-biotin. These FP-biotinylated proteins were also identified in mouse plasma by comparing band patterns on nondenaturing gels stained for albumin and carboxylesterase activity, with band patterns on blots hybridized with Streptavidin Alexa-680. Two additional FP-biotin targets, AChE (EC 3.1.1.7) and BChE (EC 3.1.1.8), were identified in mouse plasma by finding that enzyme activity was inhibited 50-80%. Mouse plasma contained eight additional FP-biotinylated bands whose identity has not yet been determined. In vitro experiments with human plasma showed that chlorpyrifos oxon, echothiophate, malaoxon, paraoxon, methyl paraoxon, diazoxon, diisopropylfluorophosphate, and dichlorvos competed with FP-biotin for binding to human albumin. Though experiments with purified albumin have previously shown that albumin covalently binds OP, this is the first report of OP binding to albumin in a living animal. Carboxylesterase is not a biomarker in man because humans have no carboxylesterase in blood. It is concluded that OP bound to albumin could serve as a new biomarker of OP exposure in man.

KW - Acetylcholinesterase

KW - Albumin

KW - Butyrylcholinesterase

KW - FP-biotin

KW - Organophosphate

UR - http://www.scopus.com/inward/record.url?scp=13644261755&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=13644261755&partnerID=8YFLogxK

U2 - 10.1093/toxsci/kfi023

DO - 10.1093/toxsci/kfi023

M3 - Article

VL - 83

SP - 303

EP - 312

JO - Toxicological Sciences

JF - Toxicological Sciences

SN - 1096-6080

IS - 2

ER -