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

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

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.

Keywords: ABCC2; Bacillus thuringiensis; Cry1Ac; Helicoverpa armigera.; alkaline phosphatase; aminopeptidase-N; cadherin.

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Alkaline Phosphatase / metabolism
  • Aminopeptidases / metabolism
  • Animals
  • Bacillus thuringiensis / metabolism*
  • Bacillus thuringiensis Toxins
  • Bacterial Proteins / metabolism*
  • Chromatography, Liquid
  • Endotoxins / metabolism*
  • Hemolysin Proteins / metabolism*
  • Moths / microbiology*
  • Multidrug Resistance-Associated Protein 2
  • Multidrug Resistance-Associated Proteins / metabolism*
  • Protein Binding
  • Tandem Mass Spectrometry

Substances

  • Bacillus thuringiensis Toxins
  • Bacterial Proteins
  • Endotoxins
  • Hemolysin Proteins
  • Multidrug Resistance-Associated Protein 2
  • Multidrug Resistance-Associated Proteins
  • insecticidal crystal protein, Bacillus Thuringiensis
  • Alkaline Phosphatase
  • Aminopeptidases
  • Adenosine Triphosphatases

Associated data

  • GENBANK/ADH16744
  • GENBANK/GQ332571.1