Michaut L, Fehlbaum P, Moniatte M, Van Dorsselaer A, Reichhart J M, Bulet P
Institut de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Réponse Immunitaire et Développement chezles Insectes, Strasbourg, France.
FEBS Lett. 1996 Oct 14;395(1):6-10. doi: 10.1016/0014-5793(96)00992-1.
Drosomycin is a 44-residue antifungal peptide with four intramolecular disulfide bridges which have been isolated from immune-challenged Drosophila. To produce adequate amounts of this peptide for 3D-structure analysis, studies on the mode of action and activity spectrum, we expressed a synthetic cDNA in Saccharomyces cerevisiae. For this purpose, we used the mating factor alpha gene and concomitantly overexpressed the KEX2 gene to increase the yield of fully processed drosomycin. Using a combination of Edman degradation and mass spectrometry, we show that drosomycin shares the same array of intramolecular disulfide bridges than plant defensins, in addition to their sequence similarities.
果蝇霉素是一种由44个氨基酸残基组成的抗真菌肽，具有四个分子内二硫键，是从受到免疫挑战的果蝇中分离出来的。为了生产足够量的这种肽用于三维结构分析、作用模式和活性谱研究，我们在酿酒酵母中表达了一个合成cDNA。为此，我们使用了交配因子α基因，并同时过表达KEX2基因以提高完全加工的果蝇霉素的产量。通过结合埃德曼降解法和质谱分析，我们发现果蝇霉素除了与植物防御素具有序列相似性外，还具有相同的分子内二硫键排列。
