P-glycoprotein ATPase from the resistant pest, Helicoverpa armigera: Purification, characterization and effect of various insecticides on its transport function
P-glycoprotein ATPase from the resistant pest, Helicoverpa armigera: Purification, characterization and effect of various insecticides on its transport function
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DOI：
10.1016/j.bbamem.2010.02.019
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发表时间：
2010-06-01
影响因子：
3.4
通讯作者：
Sreeramulu, Kuruba
中科院分区：
文献类型：
--
作者：
Aurade, Ravindra M.;Jayalakshmi, Senigala K.;Sreeramulu, Kuruba
棉铃虫（Helicoverpa armigera）是农作物的主要害虫，对多种杀虫剂产生了抗药性。从耐除草剂的H. armigera。纯化18倍，产率3%。最适pH值和温度分别为7.4和30-40 ℃。动力学研究表明，这种酶的Km值为1.2 mM的ATP。Pgp来自H.棉铃虫的部分测序，发现是同源的哺乳动物PGPS的保守序列。农药刺激H.棉铃虫Pgp ATP酶活性的最大刺激高达40%。使用纯化的Pgp的固有色氨酸荧光的猝灭来定量杀虫剂结合。使用高亲和力的荧光底物，四甲基玫瑰胺，运输进行了监测，在真实的时间在含H。棉铃虫Pgp的存在可能是该害虫产生抗药性的原因之一。(C)2010 Elsevier B. V.保留所有权利。
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.