Characterization of a Cry1Ac-receptor alkaline phosphatase in susceptible and resistant Heliothis virescens larvae.
We reported previously a direct correlation between reduced soybean agglutinin binding to 63- and 68-kDa midgut glycoproteins and resistance to Cry1Ac toxin from Bacillus thuringiensis in the tobacco budworm (Heliothis virescens). In the present work we describe the identification of the 68-kDa glycoprotein as a membrane-bound form of alkaline phosphatase we term HvALP. Lectin blot analysis of HvALP revealed the existence of N-linked oligosaccharides containing terminal N-acetylgalactosamine required for [125I]Cry1Ac binding in ligand blots. Based on immunoblotting and alkaline phosphatase activity detection, reduced soybean agglutinin binding to HvALP from Cry1Ac resistant larvae of the H. virescens YHD2 strain was attributable to reduced amounts of HvALP in resistant larvae. Quantification of specific alkaline phosphatase activity in brush border membrane proteins from susceptible (YDK and F1 generation from backcrosses) and YHD2 H. virescens larvae confirmed the observation of reduced HvALP levels. We propose HvALP as a Cry1Ac binding protein that is present at reduced levels in brush border membrane vesicles from YHD2 larvae.
Jurat-Fuentes
Juan L
JL
Department of Entomology, University of Georgia, Athens, GA 30602-2603, USA.
Adang
Michael J
MJ
D000469:Alkaline Phosphatase / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001413:Bacillus thuringiensis / Q000502:physiology*; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001427:Bacterial Toxins / Q000737:chemistry / Q000378:metabolism*; D004198:Disease Susceptibility; D004731:Endotoxins / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D006460:Hemolysin Proteins; D007814:Larva / Q000201:enzymology* / Q000382:microbiology* / Q000502:physiology; D037102:Lectins / Q000378:metabolism; D007915:Lepidoptera / Q000201:enzymology* / Q000382:microbiology / Q000502:physiology*; D008970:Molecular Weight; D011134:Polysaccharides / Q000378:metabolism
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004731:Endotoxins; D006460:Hemolysin Proteins; D037102:Lectins; D011134:Polysaccharides; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D000469:Alkaline Phosphatase