Journal Title
Title of Journal: Cell Biol Toxicol
|
Abbravation: Cell Biology and Toxicology
|
Publisher
Springer Netherlands
|
|
|
|
|
|
Authors: XinKui Fang DanFeng Huang ZhaoXia Wang ChunLing Wan Tao Sun WangJie Xu ChunYan Liu Pei Zhou ZhongDong Qiao
Publish Date: 2007/05/07
Volume: 23, Issue: 6, Pages: 445-
Abstract
In order to reveal the metabolic reaction to the presence of fenvalerate mediated by P450 in insects we used the trypan blue exclusion technique and 345dimethylthiazol25diphenyltrazolium bromide MTT reduction assay to assess the vitality of Trichoplusia ni Tn cells treated with fenvalerate and observed dose and timedependent changes in total cellular P450s In addition twodimensional electrophoresis 2DE and matrixassisted laser desorption ionization timeofflight mass spectrometry MALDITOFMS were used to identify the proteins involved in the fenvalerate reaction process Finally the cDNA of P450 fragments was cloned and realtime RTPCR was performed Our data showed that at the 0–15 μmol/L challenge concentration of fenvalerate at which the vitality of Tn cells was not affected p 005 there was a tendency toward a dose and timeresponse of total cellular P450s which peaked at the 9 h p 005 and 12 h p 001 time points following 125 μmol/L stimulation with fenvalerate The 2DE assay detected more than 1300 protein spots in each twodimensional gel of which 33 spots displayed significant differences Among the changed spots three isoforms of P450 were identified One of the three P450 cDNA fragments CYP4L4 was cloned and sequenced and its expression in treated Tn cells increased significantly p 001 It was found that fenvalerate induced the expression of P450s in insect cells This suggests that fenvalerate could be metabolized by CYP4L4 through a hydroxylation reaction in insect cells
Keywords:
.
|
Other Papers In This Journal:
|