Skip to main content

Advertisement

SpringerLink
Log in

Purification and expression of glutathione S-transferase from a Malaysian population of Aedes albopictus (Diptera: Culicidae)

  • Section Cellular and Molecular Biology
  • Published:
Biologia Aims and scope Submit manuscript

Abstract

Glutathione S-transferase (GST) from the 4th instar larvae of the dengue vector Aedes albopictus was purified by glutathione-agarose affinity chromatography and characterised using SDS-PAGE. The expression of the purified enzyme in the life stages and insecticide treated populations of Ae. albopictus as well as its cross-reactivity with larval GST of two dipteran species Aedes aegypti and Batrocera papayae were observed using western blotting. The purified GST had a specific activity of 196.0 ± 11 μmol/min/mg with a purification fold and yield of 28 and 69%, respectively. The SDS-PAGE analysis of the purified GST depicted a single band size of 23 kDa. The GST was expressed in all the larval and adult stages of Ae. albopictus with the exception of the pupal stage. However, the expression level in the adult stage was visibly reduced as compared to the larval stages. Western blotting analysis showed no cross-reactivity with the GST of Ae. aegypti (4th instar) and B. papayae (3rd instar) larvae. The expression of this enzyme was not inducible by exposure to the insecticides dichlorodiphenyltrichloroethane (1.25 mg/L) and malathion (0.3125 mg/L).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BSA:

bovine serum albumin

CI:

confidence interval

DDT:

dichlorodiphenyltrichloroethane

GSH:

reduced glutathione

GST:

glutathione S-transferase

LC50 :

median lethal concentration

LC90 :

lethal concentration to 90% of test population

SDS-PAGE:

sodium dodecyl salt-polyacrylamide gel electrophoresis

SE:

standard error

TBS:

Tris-buffered saline

TTBS:

10X Tris-buffered saline with 0.1% Tween 20

References

  • Abbott W.S. 1987. A method of computing the effectiveness of an insecticide. 1925. J. Am. Mosq. Control Assoc. 3: 302–303.

    CAS  PubMed  Google Scholar 

  • Blanchette B., Feng X. & Singh B.R. 2007. Marine glutathione S-transferases. Mar. Biotechnol. 9: 513–542.

    Article  CAS  PubMed  Google Scholar 

  • Chiang F.M. & Sun C.N. 1993. Glutathione transferase isozymes of diamondback moth larvae and their role in the degradation of some organophosphorus insecticides. Pestic. Biochem. Physiol. 45: 7–14.

    Article  CAS  Google Scholar 

  • Cnubben N.H.P., Rietjens I.M.C.M., Wortelboer H., van Zanden J. & van Bladeren P.J. 2001. The interplay of glutathionerelated processes in antioxidant defense. Environ. Toxicol. Pharmacol. 10: 141–152.

    Article  CAS  PubMed  Google Scholar 

  • David J.P., Strode C., Vontas J., Nikou D., Vaughan A., Pignatelli P.M., Louis C., Hemingway J. & Ranson H. 2005. The Anopheles gambiae detoxification chip: a highly specific microarray to study metabolic-based insecticide resistance in malaria vectors. Proc. Natl. Acad. Sci. USA 102: 4080–4084.

    Article  CAS  PubMed  Google Scholar 

  • Deen J.L. 2004. Editorial: the challenge of dengue vaccine development and introduction. Trop. Med. Int. Health 9: 1–3.

    Article  PubMed  Google Scholar 

  • Dou W., Wu S., Hassan M.W. & Wang, J.J. 2009. Purification and biochemical characterization of glutathione Stransferases from three strains of Liposcelis bostrychophila Badonnel (Psocoptera: Liposcelididae): implication of insecticide resistance. Pestic. Biochem. Physiol. 94: 10–14.

    Article  CAS  Google Scholar 

  • Enayati A.A., Ranson H. & Hemingway J. 2005. Insect glutathione transferases and insecticide resistance. Insect Mol. Biol. 14: 3–8.

    Article  CAS  PubMed  Google Scholar 

  • Grant D.F., Dietze E.C. & Hammock B.D. 1991. Glutathione S-transferase isoenzyme in Aedes aegypti: purification, characterization, and isozyme-specific regulation. Insect Biochem. 21: 421–433.

    Article  CAS  Google Scholar 

  • Guzmán M.G., Muné M. & Kourí G. 2004. Dengue vaccine: priorities and progress. Expert Rev. Anti-infect. Ther. 2: 895–911.

    Article  PubMed  Google Scholar 

  • Habig W.H., Pabst M.J. & Jakoby W.B. 1974. Glutathione Stransferases: the first enzymatic step in mercapturic acid formation. J. Biol. Chem. 249: 7130–7139.

    CAS  PubMed  Google Scholar 

  • Harlow E. & Lane D. 1988. Adjuvants, pp. 96–124. In: Harlow E. & Lane D. (eds), Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.

    Google Scholar 

  • Hayes J.D., Flanagan J.U. & Jowsey I.R. 2005. Glutathione transferases. Annu. Rev. Pharmacol. Toxicol. 45: 51–88.

    Article  CAS  PubMed  Google Scholar 

  • Hedrick J.L. & Smith A.J. 1968. Size and charge isomer separation and estimation of molecular weights of proteins by disc gel electrophoresis. Arch. Biochem. Biophys. 126: 155–164.

    Article  CAS  PubMed  Google Scholar 

  • Jowett T., Wajidi M.F.F., Oxtoby E. & Wolf C.R. 1991. Mammalian genes expressed in Drosophila: a transgenic model for the study of mechanisms of chemical mutagenesis and metabolism. EMBO J. 10: 1075–1081.

    CAS  PubMed  Google Scholar 

  • Kasai S., Komagata O., Okamura Y. & Tomita T. 2009. Alternative splicing and developmental regulation of glutathione transferases in Culex quinquefasciatus Say. Pestic. Biochem. Physiol. 94: 21–29.

    Article  CAS  Google Scholar 

  • Laemmli U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685.

    Article  CAS  PubMed  Google Scholar 

  • Li X., Schuler M.A. & Berenbaum M.R. 2007. Molecular mechanisms of metabolic resistance to synthetic and natural xenobiotics. Annu. Rev. Entomol. 52: 231–253.

    Article  PubMed  Google Scholar 

  • Lowry O.H., Rosebrough N.J., Farr A.L. & Randall R.J. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193: 265–275.

    CAS  PubMed  Google Scholar 

  • Low W.Y., Feil S.C., Ng H.L., Gorman M.A., Morton C.J., Pyke J., McConville M.J., Bieri M., Mok Y.F., Robin C., Gooley P.R., Parker M.W. & Batterham P. 2010. Recognition and detoxification of the insecticide DDT by Drosophila melanogaster glutathione S-transferase D1. J. Mol. Biol. 399: 358–366.

    Article  CAS  PubMed  Google Scholar 

  • Lumjuan N., McCarroll L., Prapanthadara L.A., Hemingway J. & Ranson, H. 2005. Elevated activity of an Epsilon class glutathione transferase confers DDT resistance in the dengue vector, Aedes aegypti. Insect Biochem. Mol. Biol. 35: 861–871.

    Article  CAS  PubMed  Google Scholar 

  • Lumjuan N., Rajatileka S., Changsom D., Wicheer J., Leelapat P., Prapanthadara L.A., Somboon P., Lycett G. & Ranson H. 2011. The role of the Aedes aegypti Epsilon glutathione transferases in conferring resistance to DDT and pyrethroid insecticides. Insect Biochem. Mol. Biol. 41: 203–209.

    Article  CAS  PubMed  Google Scholar 

  • Lumjuan N., Stevenson B.J., Prapanthadara L.A., Somboon P., Brophy P.M., Loftus B.J., Severson D.W. & Ranson H. 2007. The Aedes aegypti glutathione transferase family. Insect Biochem. Mol. Biol. 37: 1026–1035.

    Article  CAS  PubMed  Google Scholar 

  • McCall P.J. & Kittayapong P. 2007. Control of dengue vectors: tools and strategies, pp. 110–119. In: UNICEF/UNDP/Worldbank/WHO (eds), Special Programme for Research & Training in Tropical Diseases (TDR), Scientific Working Group Report on Dengue, 1–5 October, 2006, Geneva, Switzerland.

    Google Scholar 

  • Oppenoorth F.J., van der Pas L.J.T. & Houx N.W.H. 1979. Glutathione S-transferase and hydrolytic activity in a tetrachlorvinphos-resistant strain of housefly and their influence on resistance. Pestic. Biochem. Physiol. 11: 176–188.

    Article  CAS  Google Scholar 

  • Öžtetik E. 2008. A tale of plant glutathione S-transferases: since 1970. Bot. Rev. 74: 419–437.

    Article  Google Scholar 

  • Prapanthadara L.A., Hemingway J. & Ketterman A.J. 1993. Partial purification and characterization of glutathione Stransferases involved in DDT resistance from the mosquito Anopheles gambiae. Pestic. Biochem. Physiol. 47: 119–133.

    Article  CAS  Google Scholar 

  • Qin G., Jia M., Liu T., Xuan T., Zhu K.Y., Guo Y., Ma E. & Zhang J. 2011. Identification and characterisation of ten glutathione S-transferase genes from oriental migratory locust, Locusta migratoria manilensis (Meyen). Pest Manag. Sci. 67: 697–704.

    Article  CAS  PubMed  Google Scholar 

  • Ranson H., Burhani J., Lumjuan N. & Black IV W.C. 2010. Insecticide resistance in dengue vectors. TropIKA.net 1: http://journal.tropika.net/pdf/tropika/v1n1/a03v1n1.pdf.

    Google Scholar 

  • Ranson H. & Hemingway J. 2005. Mosquito glutathione transferases. Methods Enzymol. 401: 226–241.

    Article  CAS  PubMed  Google Scholar 

  • Salinas A.E. & Wong M.G. 1999. Glutathione S-transferases — a review. Curr. Med. Chem. 6: 279–309.

    CAS  PubMed  Google Scholar 

  • Samra A.I., Kamita S.G., Yao H.W., Cornel A.J. & Hammock B.D. 2012. Cloning and characterization of two glutathione S-transferases from pyrethroid-resistant Culex pipiens. Pest Manag. Sci. 68: 764–772.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Towbin H., Staehelin T. & Gordon J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76: 4350–4354.

    Article  CAS  PubMed  Google Scholar 

  • Valles S.M., Perera O.P. & Strong C.A. 2003. Purification, biochemical characterization, and cDNA cloning of a glutathione S-transferase from the red imported fire ant, Solenopsis invicta. Insect Biochem. Mol. Biol. 33: 981–988.

    Article  CAS  PubMed  Google Scholar 

  • van den Berg H., Zaim M., Yadav R.S., Soares A., Ameneshewa B., Mnzava A., Hii J., Dash A.P. & Ejov M. 2012. Global trends in the use of insecticides to control vector-borne diseases. Environ. Health Perspect. 120: 577–582.

    Article  PubMed Central  PubMed  Google Scholar 

  • Vanhaelen N., Francis F. & Haubruge E. 2004. Purification and characterization of glutathione S-transferases from two syrphid flies (Syrphus ribesii and Myathropa florae). Comp. Biochem. Physiol. B Biochem. Mol. Biol. 137: 95–100.

    Article  PubMed  Google Scholar 

  • Wei S.H., Clark A.G. & Syvanen M. 2001. Identification and cloning of a key insecticide-metabolizing glutathione Stransferase (MdGST-6A) from a hyper insecticide-resistant strain of the housefly Musca domestica. Insect Biochem. Mol. Biol. 31: 1145–1153.

    Article  CAS  PubMed  Google Scholar 

  • WHO 2005. Guidelines for laboratory and field testing of mosquito larvicides, WHO/CDS/WHOPES/GCDPP/2005. 13, World Health Organization, Geneva, Switzerland. http://whqlibdoc.who.int/hq/2005/who_cds_whopes_gcdpp 2005.13.pdf.

    Google Scholar 

  • WHO 2012. Dengue and severe dengue, Fact sheet No. 117, November 2012. http://www.who.int/mediacentre/factsheets/fs117/en.

    Google Scholar 

  • Wu S., Dou W., Wu J.J. & Wang J.J. 2009. Purification and partial characterization of glutathione S-transferase from insecticide resistant field populations of Liposcelis paeta Pearman (Psocoptera: Liposcelididae). Arch. Insect Biochem. Physiol. 70: 136–150.

    Article  CAS  PubMed  Google Scholar 

  • Yamamoto K., Miake F. & Aso Y. 2007. Purification and characterization of a novel sigma-class glutathione S-transferase of the fall webworm, Hyphantria cunea. J. Appl. Entomol. 131: 466–471.

    Article  CAS  Google Scholar 

  • Zhou W.W., Li X.W., Quan Y.H., Cheng J., Zhang C.X., Gurr G. & Zhu Z.R. 2012. Identification and expression profiles of nine glutathione S-transferase genes from an important rice phloem sap-sucker and virus vector Laodelphax striatellus (Fallén) (Hemiptera: Delphacidae). Pest Manag. Sci. 68: 1296–1305.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Molecular Entomology Research Group, School of Distance Education, Universiti Sains Malaysia, 11800, Minden, Penang, Malaysia

    Syuriza A. Shukor, Mustafa F. F. Wajidi & Silas W. Avicor

  2. Vector Control Research Unit, School of Biological Sciences, Universiti Sains Malaysia, 11800, Minden, Penang, Malaysia

    Zairi Jaal

Authors
  1. Syuriza A. Shukor
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mustafa F. F. Wajidi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Silas W. Avicor
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zairi Jaal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Silas W. Avicor.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shukor, S.A., Wajidi, M.F.F., Avicor, S.W. et al. Purification and expression of glutathione S-transferase from a Malaysian population of Aedes albopictus (Diptera: Culicidae). Biologia 69, 530–535 (2014). https://doi.org/10.2478/s11756-014-0342-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11756-014-0342-3

Key words

Search

Navigation