During the past decade the pesticidal bacterium Bacillus thuringiensis has been the subject of intensive research. These efforts have yielded considerable data about the complex relationships between the structure, mechanism of action, and genetics of the organism’s pesticidal crystal proteins, and a coherent picture of these relationships is beginning to emerge. Other studies have focused on the ecological role of the B. thuringiensis crystal proteins, their performance in agricultural and other natural settings, and the evolution of resistance mechanisms in target pests. Armed with this knowledge base and with the tools of modern biotechnology, researchers are now reporting promising results in engineering more-useful toxins and formulations, in creating transgenic plants that express pesticidal activity, and in constructing integrated management strategies to insure that these products are utilized with maximum efficiency and benefit.

Bacillus thuringiensis and Its Pesticidal Crystal Proteins

Insecticidal crystal proteins of Bacillus thuringiensis.

characterized by its ability to produce crystalline inclusions during spor-ulation. TIhese inclusions consist of proteins exhibiting a highly specific insecticidal activity (reviewed in references 4 and 97). Many B. thuriuigiensis strains with different insect host spectra have been identified (9). TIhey ar-e classified into different serotypes or subspecies based on their flagellar antigens. Most strains are active against larvae of certain members of the Lepidoptera, but some show toxicity against dipterian (reviewed in refer-ence 22) or coleopteran (53) species. For several cryst.al-producing strains.

Insecticidal crystal proteins of Bacilllus thuringiensis

https://science.sciencemag.org/content/sci/177/4053/949.full.pdf

Lectins: cell-agglutinating and sugar-specific proteins.

Publisher Summary Lectins play an important role in the development of immunology. Lectins also find application in serological laboratories for typing blood and determining secretor status, separating leucocytes from erythrocytes, and agglutinating cells from blood in the preparation of plasma. They serve as reagents for the detection, isolation, and characterization of carbohydrate-containing macromolecules, including blood-group antigens. In their interaction with saccharides, lectins serve as models for carbohydrate-specific antibodies, with the important advantage to purify lectins in gram quantities. Lectins are classified according to their carbohydrate-binding specificity that includes D-mannose(D-glucose)-binding lectins and 2-acetamido-2-deoxy-D-glucose-binding lectins. The chapter considers only those lectins that have been purified to homogeneity, and studied with regard to their biophysical, biochemical, and carbohydrate-binding specificity. The chapter also describes the cell-binding and biological properties of lectins. The chapter concludes with the description of several glycopeptide structures showing the carbohydrate-binding loci with which various lectins interact.

The lectins : carbohydrate-binding proteins of plants and animals

The Position of the Active Tryptophan Residue in Lysozyme

The toxic lectin, ricin D, contains mannose, fucose, xylose, and N-acetylglucosamine as sugar components. Sugar chains are linked to Asn-10 of the A-chain, and to Asn-95 and Asn-135 of the B-chain (Funatsu, G. et al. (1978) Agric. Biol. Chem. 42, 501-503; Araki, T. & Funatsu, G. (1985) FEBS Lett. 191, 121-124). Asparagine-linked sugar chains of each glycopeptide from ricin D were liberated by hydrazinolysis followed by N-acetylation. The reducing end residues of the sugar chains were coupled with 2-aminopyridine and the pyridylamino (PA-) derivatives obtained were purified by gel-filtration and reversed-phase HPLC. Eight main PA-sugar chains were obtained from three glycopeptides and the structures of these sugar chains were determined by component analysis, stepwise exoglycosidase digestions, partial acetolysis, and 500 MHz 1H-NMR spectroscopy. The results show that oligomannose type sugar chains (Man6-7GlcNAc2) are linked to Asn-95; Man5-7 GlcNAc2 and M4X (structure, see below) to Asn-135 of the B-chain, and M3FX and M3X to Asn-10 of the A-chain. (Formula: see text).

Structures of sugar chains of ricin D.

Biochemical studies on ricin. i. purification of ricin.

Urea-denatured alpha-zein was almost completely hydrolyzed into small peptides by digestion with 1/100 (w/w) of thermolysin at 37 degrees C for 3h. The angiotensin I-converting enzyme (ACE) inhibitory activity (IC50) of the thermolysin digest of total alpha-zein was 24.5 micrograms/ml, and most of the peptide fractions from Z19 alpha-zein and total alpha-zein separated by reverse-phase HPLC had more or less ACE inhibitory activity. From these fractions, thirty-six peptides, including 5 dipeptides, 14 tripeptides, 9 tetrapeptides, 5 pentapeptides, and 3 hexapeptides, were purified and their amino acid sequences were determined.

Isolation from α-Zein of Thermolysin Peptides with Angiotensin I-Converting Enzyme Inhibitory Activity

Purified crystal protein from Bacillus thuringiensis subsp. dendrolimus strain T84A1 gave a single polypeptide band (apparent mol. wt. 145,000) on SDS-PAGE and the only N-terminal amino acid sequence was Met–Asp–Asn–Asn–Pro–Asn–Ile–. No pentose or hexose was detected. Solid phase tryptic digestion of the solubilized crystal protein bound on a DEAE-cellulose column was found to be a successful method to prepare a toxic fragment without denaturation. The resulting fragment (fragment T) was slightly basic (pI 8.9) and the molecular weight was estimated as 58,000 on SDS-PAGE and by gel filtration without denaturing agents. The apparent number of the amono acid residues was 516 and the N- and C-terminal sequences were determined as Ile–Glu–X–Gly–Tyr–Thr– and –Leu–Arg, respectively. The molar specific toxicities of the solubilized crystal protein and fragment T were nearly identical. The fragment T retained the full toxicity and selectivity of the intat crystal protein. Location of the fragment T in the crystal...

Gunki Funatsu

Papers

The Position of the Active Tryptophan Residue in Lysozyme

Structures of sugar chains of ricin D.

Biochemical studies on ricin. i. purification of ricin.

Isolation from α-Zein of Thermolysin Peptides with Angiotensin I-Converting Enzyme Inhibitory Activity

A Toxic Fragment from the Entomocidal Crystal Protein of Bacillus thuringiensis