Glycolipids as Receptors for Bacillus thuringiensis Crystal Toxin

Science ◽  
2005 ◽  
Vol 307 (5711) ◽  
pp. 922-925 ◽  
Author(s):  
J. S. Griffitts
Keyword(s):  
Bacillus Thuringiensis ◽  
Crystal Toxin

scholarly journals The Drosophila melanogaster homologue of the human histo-blood group Pk gene encodes a glycolipid-modifying α1,4-N-acetylgalactosaminyltransferase

2004 ◽  
Vol 382 (1) ◽  
pp. 67-74 ◽  
Author(s):  
Ján MUCHA ◽  
Jiří DOMLATIL ◽  
Günter LOCHNIT ◽  
Dubravko RENDIĆ ◽  
Katharina PASCHINGER ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Bacillus Thuringiensis ◽  
Pichia Pastoris ◽  
Genetic Basis ◽  
Methylation Analysis ◽  
Yeast Pichia Pastoris ◽  
Glycan Receptors ◽  
Cloned Cdna ◽  
Crystal Toxin ◽  
Culture Supernatants

Insects express arthro-series glycosphingolipids, which contain an α1,4-linked GalNAc residue. To determine the genetic basis for this linkage, we cloned a cDNA (CG17223) from Drosophila melanogaster encoding a protein with homology to mammalian α1,4-glycosyltransferases and expressed it in the yeast Pichia pastoris. Culture supernatants from the transformed yeast were found to display a novel UDP-GalNAc:GalNAcβ1,4GlcNAcβ1-R α-N-acetylgalactosaminyltransferase activity when using either a glycolipid, p-nitrophenylglycoside or an N-glycan carrying one or two terminal β-N-acetylgalactosamine residues. NMR and MS in combination with glycosidase digestion and methylation analysis indicate that the cloned cDNA encodes an α1,4-N-acetylgalactosaminyltransferase. We hypothesize that this enzyme and its orthologues in other insects are required for the biosynthesis of the N5a and subsequent members of the arthro-series of glycolipids as well as of N-glycan receptors for Bacillus thuringiensis crystal toxin Cry1Ac.

Alkaline extraction of toxin from spores of the mosquito pathogen, Bacillus sphaericus strain 1593

1983 ◽  
Vol 29 (2) ◽  
pp. 271-275 ◽  
Author(s):  
Elizabeth West Davidson
Keyword(s):  
Molecular Weight ◽  
Bacillus Thuringiensis ◽  
Bacillus Sphaericus ◽  
Alkaline Extraction ◽  
Toxic Activity ◽  
Crystal Toxin ◽  
Mosquito Pathogen

Toxin was extracted from spores of the mosquito pathogen Bacillus sphaericus strain 1593 using 0.05 M NaOH. The molecular weight of this toxin was 35 000–54 000. Toxic activity of this extract was resistant to a variety of enzymes including subtilisin, but was degraded by pronase. Antiserum produced to 1593 spore toxin neutralized spore toxin and cytoplasmic toxin activity, but did not react with Bacillus thuringiensis var. israelensis crystal toxin, nor did var. israelensis toxin antiserum react with B. sphaericus toxin. Crystallike parasporal inclusions accompanying the B. sphaericus 1593 spores were removed by NaOH extraction.

CHARACTERIZATION OF CULTURED INSECT CELLS SELECTED BY BACILLUS THURINGIENSIS CRYSTAL TOXIN

2004 ◽  
Vol 40 (10) ◽  
pp. 312 ◽  
Author(s):  
KAIYU LIU ◽  
BINGLIAN ZHENG ◽  
HUAZHU HONG ◽  
CAIFU JIANG ◽  
RONG PENG ◽  
...  
Keyword(s):  
Bacillus Thuringiensis ◽  
Insect Cells ◽  
Crystal Toxin

scholarly journals Purification of a protein from Bacillus thuringiensis toxic to larvae of lepidoptera

1968 ◽  
Vol 106 (2) ◽  
pp. 445-454 ◽  
Author(s):  
K. E. Cooksey
Keyword(s):  
Bacillus Thuringiensis ◽  
Gel Filtration ◽  
Pieris Brassicae ◽  
Digestion Product ◽  
Toxic Protein ◽  
Protein Toxin ◽  
Ammonium Sulphate Precipitation ◽  
Alkaline Conditions ◽  
Crystal Toxin

The protein toxin of the parasporal body or crystal of Bacillus thuringiensis (Mattés isolate) has been purified severalfold by a combination of Sephadex G-200 gel filtration and ammonium sulphate precipitation. It has been shown that the use of highly alkaline conditions for dissolution of the crystals does not lead to serious artifacts. The crystal toxin has been shown to be quantitatively related to the crystal antigen. It is possible that there is a second distinct toxin present in the crystal and this too can be detected by its antigenic reaction. Purified toxic protein has been hydrolysed in vitro by regurgitated Pieris brassicae gut enzymes, chymotrypsin, trypsin and subtilisin. In each case the digest contained a product that was still antigenic, had mol.wt. about 40000 and was toxic to P. brassicae larvae. Smaller toxic molecules (mol.wt. approx. 10000) that did not react as antigens were also produced by proteolysis. It is possible that these smaller molecules were hydrolytic products of the larger digestion product.

Inhibition of Bacillus thuringiensis proteases and their effects on crystal toxin proteins and cell-free translations

1988 ◽  
Vol 34 (6) ◽  
pp. 740-747 ◽  
Author(s):  
Margaret Bibilos ◽  
Robert E. Andrews Jr.
Keyword(s):  
Molecular Weight ◽  
Bacillus Thuringiensis ◽  
Proteolytic Activity ◽  
Exponential Phase ◽  
Late Exponential Phase ◽  
Stages Of Growth ◽  
Translational Activity ◽  
Low Levels ◽  
Crystal Toxin ◽  
Neutral Metal

Proteases produced during growth and sporulation of four strains of Bacillus thuringiensis were examined. Low levels of proteolytic activity were detected during the late exponential phase of growth in all four strains: two strains of B. thuringiensis subsp. kurstaki and one strain each of subsp. israelensis and berliner. In all strains, protease activities increased dramatically at the onset of sporulation. The principal proteases, both extra- and intra-cellular, were neutral, metal-loproteases. The pH optima and substrate specificities of proteases extracted from cells in various stages of growth and sporulation indicated that substantial diversity existed among the strains. Intracellular proteases from all four strains converted the 135 000 molecular weight protoxin of strain HD251, an isolate previously shown to have reduced intracellular proteolytic activity and which normally does not contain a protein of the lower molecular weight, to a 68 000 molecular weight toxin. Cell-free translational activity of extracts from strain HD251 were approximately fivefold more active than were equivalent extracts from strain LB1 presumably because of the reduced intracellular proteolytic activity of this strain.

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