[en] Isoenzymes of glutathione S-transferase (GST) in adult Adalia bipunctata, an aphidophagous predator, were studied. Cytosolic GST activity was studied in each beetle developmental stage. The highest activities towards both 1-chloro-2,4-dinitrobenzene (CDNB) and 2,4-dinitro-1-iodobenzene (DNIB) occurred in adults. The enzyme distribution was investigated in adults. While most of the enzymatic activity was found in the abdomen (40-50 and 34-63% respectively) using several concentrations of both CDNB and DNIB, significant differences were observed for the head and the thorax depending on the substrate. Activities were more abundant in the thorax with DNIB (37-47%) compared to the 13-19% obtained with CDNB. Some GST activity was also detected in the elytra. GSTs were purified by epoxy-activated Sepharose 6B affinity chromatography and applied to an HPLC column to determine the native molecular weight (69 kDa). Three isoenzymes were separated by chromatofocusing at pH ranges 7-4. Three bands with molecular mass from 23 to 26 kDa were visualised on SDS-PAGE. Their isoelectric points were 6.66, 6.36, and 6.21. The substrate specificities and the kinetic parameters (Vm and Km) of the isoenzymes showed large differences depending on the isoenzyme. Arch.
Bogaards JJP, Van Ommen B, Falke HE, Willems MI, Van Bladeren PJ. 1990. Glutathione S-transferases subunit induction patterns of Brussels sprouts, allyl isothiocyanate and goitrin in rat liver and small intestinal mucosa: a new approach for the identification of inducing xenobiotics. Fed Chem Toxicol 28:81-88.
Boyland E, Chasseaud LF. 1969. The role of glutathione and glutathione S-transferases in mercapturic acid biosynthesis. Adv Enzymol 32:173-219.
Clark AG. 1990. The glutathione S-transferases and resistance to insecticides. In: Hayes JD, Pickett CB, Mantle TJ, editors. Glutathione S-transferases and drug resistance. London: Taylor & Francis. p 369-379.
Clark AG, Shamaan NA, Dauterman WC, Hayaoka T. 1984. Characterization of multiple glutathione S-transferases from the house fly, Musca domestica (L.). Pest Biochem Physiol 22:51-59.
Dauterman WC. 1989. The role of glutathione S-transferases in herbicide tolerance and resistance. In: Hayes JD, Pickett CB, Mantle TJ, editors. Glutathione S-transferases and drug resistance. London: Taylor & Francis. p 347-357.
Doctor JS, Fristrom JW. 1985. Macromolecular changes in imaginal discs during postembryonic development. In: Kerkut GA, Gilbert LI, editors. Comprehensive insect physiology biochemistry and pharmacology, postembryonic development. Oxford: Pergamon. p 201-238.
Egaas E, Svendsen NO, Kobro S, Skaare JU, Jensen EG. 1991. Activities and properties of xenobiotic metabolizing enzymes in the peach-potato aphid (Myzus persicae Sultzer) feeding on paprika (Capsicum annuun L.) or swedes (Brassica napus rapifera Metzer). Comp Biochem Physiol 99: 105-110.
Fournier D, Bride JM, Poirié M, Bergé JB, Plapp FW. 1992. Insect glutathione S-transferases. Biochemical characteristics of the major forms from houseflies susceptible and resistant to insecticides. J Biol Chem 267:1840-1845.
Francis F. 1999. Conséquences évolutives des relations entre le puceron et son préédateur en présence de substances allélochimiques chez les Brassicacae. Proc 5th Conf Intern Rav Agr 2:503-510.
Francis F, Haubruge E, Gaspar C. 1999. Effects of isothiocyanates on the glutathione S-transferases activity from Adalia bipunctata L. (Coleoptera:Coccinellidae). Med Fac Landbouw Univ Gent 64:297-303.
Francis F, Lognay G, Wathelet JP, Haubruge E. 2001. Effects of allelochemicals from first (Brassicaceae) and second (Myzus persicae & Brevicoryne brassicae) trophic levels on Adalia bipunctata. J Chem Ecol 27:243-256.
Gillott C. 1980. Entomology. New York: Plenum Press, 386 p.
Grant DF, Matsumura F. 1989. Glutathione S-transferase 1 and 2 in susceptible and insecticide resistant Aedes aegypti. Pestic Biochem Physiol 33:132-143.
Habig WH, Pabst MJ, Jakoby WB. 1974. Glutathione S-transferases: the first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130-7139.
Hemingway J, Malcolm CA, Kissoon KE, Boddington RG, Curtis CF, Hill N. 1985. The biochemistry of insecticide resistance in Anopheles sacharovi: comparative studies with a range of insecticide susceptible and resistant Anopheles and Culex species. Pest Biochem Physiol 24:68-76.
Konno Y, Shishido T. 1992. Glutathione S-transferase distribution in insect tissues. Appl Ent Zool 27:391-397.
Kostaropoulos I, Papadopoulos AI. 1998. Glutathione S-transferase isoenzymes expressed in the three developmental stages of the insect Tenebrio molitor. Insect Biochem Mol Biol 28:901-909.
Kostaropoulos I, Mantzari AE, Papadopoulos AI. 1996. Alterations of some glutathione S-transferase characteristics during the development of Tenebrio molitor (Insecta: Coleoptera). Insect Biochem Mol Biol 26:963-969.
Laemmli UK. 1970. Cleavage of structural proteins during assembly of head bacteriophage T4. Nature 227:680-685.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. 1951. Protein measurement with the folin phenol reagent. J Biol Chem 193:265-275.
Pickett CB, Lu AYH. 1989. Glutathione S-transferases: gene structure, regulation and biological function. Ann Rev Biochem 58:743-764.
Prapanthadara L, Promtet N, Koottathep S, Somboon P, Ketterman AJ. 2000. Isoenzymes of glutathione S-transferase from the mosquito Anopheles dirus species B: the purification, partial characterization and interaction with various insecticides. Insect Biochem Mol Biol 30: 395-403.
Saleh MA, Motoyama N, Dauterman WC. 1978. Reduced glutathione in the housefly: concentration during development and variation in strains. Insect Biochem 8:311-316.
Simons PC, Vander jagt DL. 1977. Purification of glutathione S-transferases from human liver by glutathione affinity chromatography. Anal Biochem 82:334-341.
Usui K, Kukami J, Shishido T. 1977. Insect glutathione S-transferase: separation of transferases from fat bodies of American cockroaches active on organophosphorous trimesters. Pest Biochem Physiol 7:249-254.
Wadleigh R, Yu SJ, 1988. Detoxification of isothiocyanate allelochemicals by glutathione transferase in three lepidopterous species. J Chem Ecol 14:1279-1288.
Yu SJ. 1982. Host plant induction of glutathione S-transferases in the fall armyworm. Pest Biochem Physiol 19:101-106.
Yu SJ. 1987. Biochemical defence capacity in the spined soldier bug (Posidus maculinventris) and its lepidopterous prey. Pest Biochem Physiol 28:216-222.
Yu SJ. 1989. Purification and characterization of glutathione S-transferases from five phytophagous Lepidoptera. Pest Biochem Physiol 35:97-105.
Yu SJ. 1999. Induction of new glutathione S-transferase isozymes by allelochemicals in the fall armyworm. Pest Biochem Physiol 63:163-171.
Yu SJ, Hsu EL. 1993. Induction of detoxification enzymes in phytophagous insects: roles of insecticide synergists, larval age and species. Arch Insect Biochem Physiol 24:21-32.
