[32] Preparation of microsomes from insects and purification of CYP6D1 from house flies

Scott, Jeffrey G.

doi:10.1016/s0076-6879(96)72034-7

Cited by 4 publications

(1 citation statement)

References 10 publications

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“…Based on the method of Scott (1998), microsomes were thawed and 10% CHAPS in water was added with brief shaking to a final concentration of 0.5%, left on ice for up to 4 hours, then 40% (w/v) polyethylene glycol 6000 (PEG) was added to a final concentration of 10% and was incubated on ice for 30 minutes. Precipitates were removed by centrifuging at 10 000 g for 10 minutes and supernatant to a final concentration of 2 mol/L and insoluble material was pelleted at 10 000 g for 10 minutes.…”

Section: Approachesmentioning

confidence: 99%

Partial purification of Cytochrome P450 from Helicoverpa armigera (Hubner)

Gao

2005

Insect Science

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The cytochrome P450 (Cyt-P450) proteins from the fat body and midgut of the cotton bollworm, Helicoverpa armigera, were respectively partially purified by a set of purification procedures including differential centrifugation, solubilization of CHAPS, protein precipitation by PEG precipitation and DE-32 column chromatography. The Cyt-P450 was detected by methods of CO difference spectrum and SDS-PAGE. Fraction of detergent solubilized microsomes from the fat body of H. armigera was purified more than 17-fold. Three protein bands were detected by SDS-PAGE with molecular masses of 70 600, 63 300 and 571 200Da. It is possible that the proteins with molecular mass of 63 300 and 571 200Da were the isozymes of Cyt-P450.

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Section: Approachesmentioning

confidence: 99%

Partial purification of Cytochrome P450 from Helicoverpa armigera (Hubner)

Gao

2005

Insect Science

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Profiling of abundant proteins associated with dichlorodiphenyltrichloroethane resistance in Drosophila melanogaster

et al. 2005

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Dichlorodiphenyltrichloroethane (DDT) metabolism-based resistance in Drosophila melanogaster is a complex metabolic system associated with the transcription of detoxification related genes, ion transport, lipid and sugar metabolism pathways. However, little is known about the differences regarding the proteome of field- and laboratory-selected resistant Drosophila genotypes. We investigated the impact of DDT resistance in the abundant proteome of field- and laboratory- selected resistant Drosophila using a two-dimensional gel electrophoresis DDT reference map. Proteomic profiling was performed in two DDT susceptible genotypes (Canton-S and 91-C) and three DDT resistant lines (Rst(2)DDT(91-R), Rst(2)DDT(Wisconsin) and Rst(2)DDT(Hikone-R)). Protein spots were stained with Coomassie blue and compared using PDQuest software. Selected protein spots were cut out and analyzed using matrix assisted laser desorption-time of flight mass spectrometry. Querying the NCBInr. 10.21.2003 database with mass spectrometric data yielded the identity of 21 differentially translated proteins in Rst(2)DDT(91-R), Rst(2)DDT(Wisconsin) and Canton-S representing proteins putatively involved in biochemical pathways such as glycolysis and gluconeogenesis, the pentose phosphate pathway, the Krebs cycle and fatty acid oxidation. We hypothesize that both strategies are aimed to use of the pentose phosphate pathway to increase glucose utilization while Rst(2)DDT(91-R) relies primarily on glycolysis to produce reduced NADP and increase DDT detoxification. DDT exposure in Canton-S induced six proteins, while four proteins were repressed in Rst(2)DDT(Hikone-R). Our data suggest that insecticide resistance appears to impact different metabolic pathways in Drosophila genotypes selected with the same pesticide (DDT).

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New approaches to old problems: Removal of phospholipase A2 results in highly active microsomal membranes from the honey bee, Apis mellifera

Zaworra

Nauen

2019

Pesticide Biochemistry and Physiology

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[32] Preparation of microsomes from insects and purification of CYP6D1 from house flies

Cited by 4 publications

References 10 publications

Partial purification of Cytochrome P450 from Helicoverpa armigera (Hubner)

Partial purification of Cytochrome P450 from Helicoverpa armigera (Hubner)

Profiling of abundant proteins associated with dichlorodiphenyltrichloroethane resistance in Drosophila melanogaster

New approaches to old problems: Removal of phospholipase A2 results in highly active microsomal membranes from the honey bee, Apis mellifera

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