Lepage P, Bitsch F, Roecklin D, Keppi E, Dimarcq J L, Reichhart J M, Hoffmann J A, Roitsch C, Van Dorseelaer A
Transgène S. A., Strasbourg, France.
Eur J Biochem. 1991 Mar 28;196(3):735-42. doi: 10.1111/j.1432-1033.1991.tb15872.x.
The primary-structure comparison of natural insect defensin A from Phormia terranovae and recombinant insect defensin A from Saccharomyces cerevisiae has been accomplished using a combination of Edman degradation and liquid secondary ion mass spectrometry. The natural and recombinant proteins have the same primary structure with identical disulfide-bond designations (formula; see text) as determined from the peptides obtained after thermolysin digestion. The combined use of Edman degradation and mass spectometry allowed the disulfide-bridge structure to be determined with a total of only 40 micrograms (9.9 nmol) natural peptide. Mass spectrometry provides a rapid means of disulfide-bridge verification, requiring not more than 20 micrograms recombinant insect defensin A, which is compatible with use in batch analysis.
已通过埃德曼降解法和液体二次离子质谱法相结合的方式，完成了对来自新大陆绿蝇的天然昆虫防御素A与来自酿酒酵母的重组昆虫防御素A的一级结构比较。天然蛋白和重组蛋白具有相同的一级结构，且二硫键的命名相同（分子式；见正文），这是根据嗜热菌蛋白酶消化后得到的肽段确定的。埃德曼降解法和质谱法的联合使用，仅用总共40微克（9.9纳摩尔）的天然肽段，就确定了二硫桥结构。质谱法提供了一种快速验证二硫桥的方法，所需的重组昆虫防御素A不超过20微克，适用于批量分析。
