P-glycoprotein ATPase from the resistant pest, Helicoverpa armigera: Purification, characterization and effect of various insecticides on its transport function

被引：53

作者：

Aurade, Ravindra M. ^{[1
]}

Jayalakshmi, Senigala K. ^{[2
]}

Sreeramulu, Kuruba ^{[1
]}

机构：

[1] Gulbarga Univ, Dept Biochem, Gulbarga 585106, India

[2] Univ Agr Sci, Agr Res Stn, Gulbarga 585103, India

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES | 2010年 / 1798卷 / 06期

关键词：

Helicoverpa armigera; P-glycoprotein ATPase; Proteoliposomes; Insecticides; Tryptophan quenching; Drug transport; HAMSTER OVARY CELLS; MULTIDRUG-RESISTANCE; TRANSEPITHELIAL TRANSPORT; MALPIGHIAN TUBULES; TOBACCO HORNWORM; DRUG TRANSPORT; BINDING; ABC; MECHANISM; NUCLEOTIDE;

D O I：

10.1016/j.bbamem.2010.02.019

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Helicoverpa armigera is a major pest of agricultural crops and has developed resistance to various insecticides. A P-glycoprotein (Pgp) with ATPase activity likely to be involved in insecticide resistance was purified and characterized from insecticide-resistant H. armigera. The purification was 18-fold with 3% yield. The optimum pH and temperature were found to be 7.4 and 30-40 degrees C, respectively. Kinetic studies indicated that this enzyme had a Km value of 1.2 mM for ATP. Pgp from H. armigera was partially sequenced and found to be homologous to conserved sequences of mammalian Pgps. Pesticides stimulated H. armigera Pgp ATPase activity with a maximum stimulation of up to 40%. Quenching of the intrinsic tryptophan fluorescence of purified Pgp was used to quantitate insecticide binding. Using the high-affinity fluorescent substrate, tetramethylrosamine, transport was monitored in real time in proteoliposomes containing H. armigera Pgp. The presence of Pgp could be one of the reasons for insecticide resistance in this pest. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：1135 / 1143

页数：9

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