Biochemical characterization of digestive membrane-associated alkaline phosphatase from the velvet bean caterpillar Anticarsia gemmatalis

被引:9
作者
da Silva, Gabriela [1 ]
Costa Ramos, Luis Felipe [1 ]
Seckler, Henrique dos Santos [1 ]
Gomes, Fabio Mendonca [2 ]
Cortines, Juliana Reis [3 ]
Ramos, Isabela [4 ]
Anobom, Cristiane Dinis [1 ]
Machado, Ednildo de Alcantara [2 ]
Perpetua de Oliveira, Danielle Maria [1 ]
机构
[1] Univ Fed Rio de Janeiro, Ctr Ciencias Matemat & Nat, Lab Bioquim Estrutural Prot, Dept Bioquim,Inst Quim, BR-21941909 Rio De Janeiro, Brazil
[2] Univ Fed Rio de Janeiro, Inst Biofis Carlos Chagas Filho, Lab Bioquim Insetos & Parasitos, Rio De Janeiro, Brazil
[3] Univ Fed Rio de Janeiro, Inst Microbiol Paulo Goes, Ctr Ciencias Saude, Dept Virol, Rio De Janeiro, Brazil
[4] Univ Fed Rio de Janeiro, Inst Bioquim Med, Ctr Ciencias Saude, Lab Bioquim Insetos, Rio De Janeiro, Brazil
关键词
alkaline phosphatase; Anticarsia gemmatalis; biological control; Cry1Ac; pest insect; BACILLUS-THURINGIENSIS; MANDUCA-SEXTA; MIDGUT; BINDING; CRY1AC; ISOZYMES; PROTEINS; AMINOPEPTIDASE; PURIFICATION; LEPIDOPTERA;
D O I
10.1002/arch.21591
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
In Brazil, the use of transgenic plants expressing the insect-toxic Bacillus thuringiensis endotoxin has been successfully used as pest control management since 2013 in transgenic soybean lineages against pest caterpillars such as Helicoverpa armigera. These toxins, endogenously expressed by the plants or sprayed over the crops, are ingested by the insect and bind to receptors in the midgut of these animals, resulting in disruption of digestion and lower insect survival rates. Here, we identified and characterized a membrane-associated alkaline phosphatase (ALP) in the midgut of Anticarsia gemmatalis, the main soybean defoliator pest in Brazil, and data suggested that it binds to Cry1Ac toxin in vitro. Our data showed a peak of ALP activity in homogenate samples of the midgut dissected from the 4th and 5th instars larvae. The brush border membrane vesicles obtained from the midgut of these larvae were used to purify a 60 kDa ALP, as detected by in-gel activity and in vitro biochemical characterization using pharmacological inhibitors and mass spectrometry. When Cry1Ac toxin was supplied to the diet, it was efficient in decreasing larval weight gain and survival. Indeed, in vitro incubation of Cry1Ac toxin with the purified ALP resulted in a 43% decrease in ALP specific activity and enzyme-linked immunosorbent assay showed that ALP interacts with Cry1Ac toxin in vitro, thus suggesting that ALP could function as a Cry toxin ligand. This is a first report characterizing an ALP in A. gemmatalis.
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页数:14
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