State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Chaoyang District, Beijing, China.
Mol Biotechnol. 2010 Mar;44(3):177-89. doi: 10.1007/s12033-009-9224-2.
We attempted to identify membrane proteins associated with the glycoconjugates and cell wall biosynthesis in the total membrane preparations of Aspergillus fumigatus. The total membrane preparations were first run on 1D gels, and then the stained gels were cut and submitted to in-gel digestion followed by 2D LC-MS/MS and database search. A total of 530 proteins were identified with at least two peptides detected with MS/MS spectra. Seventeen integral membrane proteins were involved in N-, O-glycosylation or GPI anchor biosynthesis. Nine membrane proteins were involved in cell wall biosynthesis. Eight proteins were identified as enzymes involved in sphingolipid synthesis. In addition, the proteins involved in cell wall and ergosterol biosynthesis can potentially be used as antifungal drug targets. Our method, for the first time, clearly provided a global view of the membrane proteins associated with glycoconjugates and cell wall biosynthesis in the total membrane proteome of A. fumigatus.
我们试图鉴定与烟曲霉总膜制剂中糖缀合物和细胞壁生物合成相关的膜蛋白。首先在一维凝胶上运行总膜制剂，然后对染色凝胶进行切割，进行胶内消化，然后进行二维 LC-MS/MS 和数据库搜索。共鉴定到 530 种蛋白质，其中至少有 2 种肽段被 MS/MS 谱检测到。17 种完整的膜蛋白参与 N-、O-糖基化或 GPI 锚生物合成。9 种膜蛋白参与细胞壁生物合成。8 种蛋白质被鉴定为参与鞘脂合成的酶。此外，参与细胞壁和麦角固醇生物合成的蛋白质可能可作为抗真菌药物靶点。我们的方法首次清楚地提供了与烟曲霉总膜蛋白组中糖缀合物和细胞壁生物合成相关的膜蛋白的全局视图。
