Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull-down assay.

Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull-down assay.
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DOI:
10.1002/mbo3.360
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发表时间:
2016-08
期刊:
影响因子:
3.4
通讯作者:
Zhang J
Zhang J
中科院分区:
生物学3区
文献类型:
--
作者:
Zhou Z;Wang Z;Liu Y;Liang G;Shu C;Song F;Zhou X;Bravo A;Soberón M;Zhang J

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采用改进的下拉法(pull-down)结合液相色谱-串联质谱(LC-MS/MS)鉴定了棉铃虫刷状缘膜囊泡中的Cry 1Ac毒素结合蛋白。根据LC-MS/MS结果,Cry 1Ac毒素可以结合六类氨肽酶-N、碱性磷酸酶、钙粘蛋白样蛋白、ATP结合盒转运蛋白亚家族C蛋白(ABCC 2)、肌动蛋白、ATP酶、聚钙蛋白以及一些先前未被表征为Cry毒素结合分子的其他蛋白质,如二肽基肽酶或羧基/胆碱酯酶和一些丝氨酸蛋白酶。这是第一次报道Cry 1Ac毒素与棉铃虫ABCC 2的直接结合。
Cry1Ac toxin‐binding proteins from Helicoverpa armigera brush border membrane vesicles were identified by an improved pull‐down method that involves coupling Cry1Ac to CNBr agarose combined with liquid chromatography–tandem mass spectrometry (LC‐MS/MS). According to the LC‐MS/MS results, Cry1Ac toxin could bind to six classes of aminopeptidase‐N, alkaline phosphatase, cadherin‐like protein, ATP‐binding cassette transporter subfamily C protein (ABCC2), actin, ATPase, polycalin, and some other proteins not previously characterized as Cry toxin‐binding molecules such as dipeptidyl peptidase or carboxyl/choline esterase and some serine proteases. This is the first report that suggests the direct binding of Cry1Ac toxin to ABCC2 in H. armigera.