通过改进的下拉分析法鉴定ABCC2为棉铃虫刷状缘膜囊泡上Cry1Ac的结合蛋白。

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

作者信息

Zhou Zishan, Wang Zeyu, Liu Yuxiao, Liang Gemei, Shu Changlong, Song Fuping, Zhou Xueping, Bravo Alejandra, Soberón Mario, Zhang Jie

机构信息

State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, No. 866 Yuhangtang Road, Xihu District, Hangzhou, 310012, China.

State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, No. 2 Yuanmingyuan West Road, Haidian District, Beijing, 100193, China.

出版信息

Microbiologyopen. 2016 Aug;5(4):659-69. doi: 10.1002/mbo3.360. Epub 2016 Apr 1.

DOI:10.1002/mbo3.360

PMID:27037552

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4985599/

Abstract

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.

摘要

通过一种改进的下拉法鉴定了棉铃虫刷状缘膜囊泡中的Cry1Ac毒素结合蛋白，该方法包括将Cry1Ac偶联到溴化氰琼脂糖上，并结合液相色谱-串联质谱（LC-MS/MS）。根据LC-MS/MS结果，Cry1Ac毒素可与六类氨肽酶-N、碱性磷酸酶、类钙黏蛋白、ATP结合盒转运蛋白C亚家族蛋白（ABCC2）、肌动蛋白、ATP酶、多钙蛋白以及其他一些以前未被鉴定为Cry毒素结合分子的蛋白质结合，如二肽基肽酶或羧基/胆碱酯酶以及一些丝氨酸蛋白酶。这是首次报道Cry1Ac毒素在棉铃虫中直接与ABCC2结合。