Partial purification and characterization of a methyl-parathion resistance-associated general esterase in Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

Xuguo Zhou; Michael E. Scharf; Gautam Sarath; Lance J. Meinke; Laurence D. Chandler; Blair D. Siegfried

doi:10.1016/j.pestbp.2003.10.003

Partial purification and characterization of a methyl-parathion resistance-associated general esterase in Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

Xuguo Zhou, Michael E. Scharf, Gautam Sarath, Lance J. Meinke, Laurence D. Chandler, Blair D. Siegfried

Entomology

Research output: Contribution to journal › Article › peer-review

22 Scopus citations

Abstract

Increased hydrolytic metabolism of organophosphate insecticides has been associated with resistance among Nebraska western corn rootworm populations. In this study, resistance-associated esterases were partially purified by differential centrifugation, ion exchange, and hydroxyapatite column chromatography, with a final purification factor of 100-fold and recovery of approximately 10%. Kinetic analysis of the partially purified enzyme indicated that the K_m of the group II esterases was identical for the two populations, although V_max was consistently threefold higher in the resistant population. A putative esterase, DvvII, was further purified to homogeneity by preparative polyacrylamide gel electrophoresis. DvvII is a monomer with a molecular weight of approximately 66kDa, although three distinct isoforms with similar pIs were evident based on isoelectric focusing gel electrophoresis. Immunoassays with the Myzus persicae E4 antiserum indicated that group II esterases from D. v. virgifera were cross-reactive and expressed at much higher titers in the resistant population relative to the susceptible counterpart. These results suggest that the resistance is likely associated with overproduction of an esterase isozyme in resistant D. v. virgifera populations.

Original language	English
Pages (from-to)	114-125
Number of pages	12
Journal	Pesticide Biochemistry and Physiology
Volume	78
Issue number	2
DOIs	https://doi.org/10.1016/j.pestbp.2003.10.003
State	Published - Feb 2004

Bibliographical note

Funding Information:
The authors thank Z.B. Mayo and David Taylor for valuable comments on an earlier draft of the manuscript. Jim Brown and Jenny Stebbing helped collect and maintain D. v. virgifera field populations. Support for this project was provided by DuPont, FMC, Zeneca, American Cyanamid, Dow Agrosciences, USDA Specific Cooperative Agreement 58-5447-6-116, USDA Initiative for Future Agricultural and Food Science Grant No. 00-52103-9699) and NIH Grant No. 1P20RR16569 from the BRIN Program of the National Center for Research Resources.

Keywords

Characterization
Diabrotica virgifera virgifera
Esterase
Insecticide resistance
Purification

ASJC Scopus subject areas

Agronomy and Crop Science
Health, Toxicology and Mutagenesis

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10.1016/j.pestbp.2003.10.003

Cite this

@article{5012d54796254aa39a74db29f728699b,

title = "Partial purification and characterization of a methyl-parathion resistance-associated general esterase in Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)",

abstract = "Increased hydrolytic metabolism of organophosphate insecticides has been associated with resistance among Nebraska western corn rootworm populations. In this study, resistance-associated esterases were partially purified by differential centrifugation, ion exchange, and hydroxyapatite column chromatography, with a final purification factor of 100-fold and recovery of approximately 10%. Kinetic analysis of the partially purified enzyme indicated that the Km of the group II esterases was identical for the two populations, although Vmax was consistently threefold higher in the resistant population. A putative esterase, DvvII, was further purified to homogeneity by preparative polyacrylamide gel electrophoresis. DvvII is a monomer with a molecular weight of approximately 66kDa, although three distinct isoforms with similar pIs were evident based on isoelectric focusing gel electrophoresis. Immunoassays with the Myzus persicae E4 antiserum indicated that group II esterases from D. v. virgifera were cross-reactive and expressed at much higher titers in the resistant population relative to the susceptible counterpart. These results suggest that the resistance is likely associated with overproduction of an esterase isozyme in resistant D. v. virgifera populations.",

keywords = "Characterization, Diabrotica virgifera virgifera, Esterase, Insecticide resistance, Purification",

author = "Xuguo Zhou and Scharf, {Michael E.} and Gautam Sarath and Meinke, {Lance J.} and Chandler, {Laurence D.} and Siegfried, {Blair D.}",

note = "Funding Information: The authors thank Z.B. Mayo and David Taylor for valuable comments on an earlier draft of the manuscript. Jim Brown and Jenny Stebbing helped collect and maintain D. v. virgifera field populations. Support for this project was provided by DuPont, FMC, Zeneca, American Cyanamid, Dow Agrosciences, USDA Specific Cooperative Agreement 58-5447-6-116, USDA Initiative for Future Agricultural and Food Science Grant No. 00-52103-9699) and NIH Grant No. 1P20RR16569 from the BRIN Program of the National Center for Research Resources.",

year = "2004",

month = feb,

doi = "10.1016/j.pestbp.2003.10.003",

language = "English",

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Partial purification and characterization of a methyl-parathion resistance-associated general esterase in Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae). / Zhou, Xuguo; Scharf, Michael E.; Sarath, Gautam et al.
In: Pesticide Biochemistry and Physiology, Vol. 78, No. 2, 02.2004, p. 114-125.

Research output: Contribution to journal › Article › peer-review

TY - JOUR

T1 - Partial purification and characterization of a methyl-parathion resistance-associated general esterase in Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

AU - Zhou, Xuguo

AU - Scharf, Michael E.

AU - Sarath, Gautam

AU - Meinke, Lance J.

AU - Chandler, Laurence D.

AU - Siegfried, Blair D.

N1 - Funding Information: The authors thank Z.B. Mayo and David Taylor for valuable comments on an earlier draft of the manuscript. Jim Brown and Jenny Stebbing helped collect and maintain D. v. virgifera field populations. Support for this project was provided by DuPont, FMC, Zeneca, American Cyanamid, Dow Agrosciences, USDA Specific Cooperative Agreement 58-5447-6-116, USDA Initiative for Future Agricultural and Food Science Grant No. 00-52103-9699) and NIH Grant No. 1P20RR16569 from the BRIN Program of the National Center for Research Resources.

PY - 2004/2

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N2 - Increased hydrolytic metabolism of organophosphate insecticides has been associated with resistance among Nebraska western corn rootworm populations. In this study, resistance-associated esterases were partially purified by differential centrifugation, ion exchange, and hydroxyapatite column chromatography, with a final purification factor of 100-fold and recovery of approximately 10%. Kinetic analysis of the partially purified enzyme indicated that the Km of the group II esterases was identical for the two populations, although Vmax was consistently threefold higher in the resistant population. A putative esterase, DvvII, was further purified to homogeneity by preparative polyacrylamide gel electrophoresis. DvvII is a monomer with a molecular weight of approximately 66kDa, although three distinct isoforms with similar pIs were evident based on isoelectric focusing gel electrophoresis. Immunoassays with the Myzus persicae E4 antiserum indicated that group II esterases from D. v. virgifera were cross-reactive and expressed at much higher titers in the resistant population relative to the susceptible counterpart. These results suggest that the resistance is likely associated with overproduction of an esterase isozyme in resistant D. v. virgifera populations.

AB - Increased hydrolytic metabolism of organophosphate insecticides has been associated with resistance among Nebraska western corn rootworm populations. In this study, resistance-associated esterases were partially purified by differential centrifugation, ion exchange, and hydroxyapatite column chromatography, with a final purification factor of 100-fold and recovery of approximately 10%. Kinetic analysis of the partially purified enzyme indicated that the Km of the group II esterases was identical for the two populations, although Vmax was consistently threefold higher in the resistant population. A putative esterase, DvvII, was further purified to homogeneity by preparative polyacrylamide gel electrophoresis. DvvII is a monomer with a molecular weight of approximately 66kDa, although three distinct isoforms with similar pIs were evident based on isoelectric focusing gel electrophoresis. Immunoassays with the Myzus persicae E4 antiserum indicated that group II esterases from D. v. virgifera were cross-reactive and expressed at much higher titers in the resistant population relative to the susceptible counterpart. These results suggest that the resistance is likely associated with overproduction of an esterase isozyme in resistant D. v. virgifera populations.

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KW - Esterase

KW - Insecticide resistance

KW - Purification

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JF - Pesticide Biochemistry and Physiology

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Partial purification and characterization of a methyl-parathion resistance-associated general esterase in Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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