Koller C N, Bauer L S, Hollingworth R M
Pesticide Research Center, Michigan State University, E. Lansing 48824.
Biochem Biophys Res Commun. 1992 Apr 30;184(2):692-9. doi: 10.1016/0006-291x(92)90645-2.
Native crystals of Bacillus thuringiensis var. san diego, a coleopteran-specific delta-endotoxin, were metabolically labelled with [35S]methionine. Specific activity was 82,000 CPM/micrograms (2.44 Ci/mmol). Using a universal buffer formulated with the same ionic strength at every pH, we determined that native crystals dissolve above pH 10 and below pH 4. At the acidic pH, the rate of solubilization was substantially slower than at the alkaline pH. Recrystallization rates for the toxin were similar regardless of solubilization conditions. The banding patterns in denatured polyacrylamide gel electrophoresis were unaffected by solubilization conditions. Toxicity was higher for soluble toxin compared to crystal toxin, but virtually identical for the acidic and alkaline produced solutions. Acid solubilization is significant because of the acidic midgut of susceptible Coleoptera.
苏云金芽孢杆菌圣地亚哥变种(一种鞘翅目特异性δ-内毒素)的天然晶体用[35S]甲硫氨酸进行代谢标记。比活为82,000 CPM/微克(2.44 Ci/毫摩尔)。使用在每个pH值下离子强度相同的通用缓冲液,我们确定天然晶体在pH值高于10和低于4时会溶解。在酸性pH值下,溶解速率比在碱性pH值下慢得多。无论溶解条件如何,毒素的重结晶速率相似。变性聚丙烯酰胺凝胶电泳中的条带模式不受溶解条件的影响。与晶体毒素相比,可溶性毒素的毒性更高,但酸性和碱性产生的溶液的毒性几乎相同。由于易感鞘翅目昆虫的中肠呈酸性,酸溶解具有重要意义。