Identification and characterization of Aedes aegypti aminopeptidase N as a putative receptor of Bacillus thuringiensis Cry11A toxin.
Identification and characterization of Aedes aegypti aminopeptidase N as a putative receptor of Bacillus thuringiensis Cry11A toxin.
复制标题
DOI：
10.1016/j.ibmb.2009.08.003
复制
发表时间：
2009-10
影响因子：
3.8
通讯作者：
Gill, Sarjeet S.
中科院分区：
文献类型：
--
作者：
Chen, Jianwu;Aimanova, Karlygash G.;Pan, Songqin;Gill, Sarjeet S.
苏云金芽孢杆菌亚种以色列蚊在世界范围内用于控制埃及伊蚊幼虫，在产孢过程中产生Cry11Aa和其他毒素。本研究采用生物素化Cry11Aa毒素和从伊蚊幼虫中肠制备的可溶刷状膜囊泡进行下拉测定。3个洗脱蛋白被鉴定为氨肽酶N (APN)，其中一个蛋白为140 kDa，命名为AaeAPN1（在VectorBase中为AAEL 012778）。该蛋白定位于幼虫后中肠上皮细胞的顶端。克隆出全长AaeAPN1，并在大肠杆菌和Sf21细胞中表达。在Sf21细胞中表达的AaeAPN1蛋白具有酶活性，具有gpi锚点，但不结合Cry11Aa。然而，截断的AaeAPN1以高亲和力结合Cry11Aa，也结合Cry11Ba，但亲和力较低。小麦胚芽凝集素可以检测到BBMV而非Sf21表达的AaeAPN1，这表明原生而Sf21表达的APN1含有n -乙酰氨基葡萄糖部分。
Bacillus thuringiensis subsp. israelensis, which is used worldwide to control Aedes aegypti larvae, produces Cry11Aa and other toxins during sporulation. In this study, pull-down assays were performed using biotinylated Cry11Aa toxin and solubilized brush border membrane vesicles prepared from midguts of Aedes larvae. Three of the eluted proteins were identified as aminopeptidease N (APN), one of which was a 140 kDa protein, named AaeAPN1 (AAEL 012778 in VectorBase). This protein localizes to the apical side of posterior midgut epithelial cells of larva. The full-length AaeAPN1 was cloned and expressed in E. coli and in Sf21 cells. AaeAPN1 protein expressed in Sf21 cells was enzymatically active, had a GPI-anchor but did not bind Cry11Aa. A truncated AaeAPN1, however, binds Cry11Aa with high affinity, and also Cry11Ba but with lower affinity. BBMV but not Sf21 expressed AaeAPN1 can be detected by wheat germ agglutinin suggesting the native but Sf21 cell expressed APN1 contains N-acetylglucosamine moieties.
登录
查看更多内容
影响因子：
--
作者：
Abdullah MA;Valaitis AP;Dean DH
通讯作者：
Dean DH
影响因子：
2.6
作者：
Gill, M;Ellar, D
通讯作者：
Ellar, D
影响因子：
3.8
作者：
Krishnamoorthy, M.;Jurat-Fuentes, J. L.;Adang, Michael J.
通讯作者：
Adang, Michael J.
DOI：
10.1152/ajprenal.00487.2005
发表时间：
2007-05-01
影响因子：
4.2
作者：
Kang'ethe, Wanyoike;Aimanova, Karlygash G.;Gill, Sarjeet S.
通讯作者：
Gill, Sarjeet S.
影响因子：
2.9
作者：
Hua, Gang;Zhang, Rui;Adang, Michael J.
通讯作者：
Adang, Michael J.