Morrissey Bethny, Downard Kevin M
School of Molecular and Microbial Biosciences, The University of Sydney, Sydney, Australia.
Proteomics. 2006 Apr;6(7):2034-41. doi: 10.1002/pmic.200500642.
A proteomics-based approach is described that combines gel electrophoresis and MS in order to identify protein interactions and the nature of the interaction interface with high-sample throughput and sensitivity. Results for protein antigens of the influenza virus have demonstrated that the approach can be successfully employed to detect determinants within the hemagglutinin antigen of two divergent type A forms of the virus in present circulation. The determinants are localised to residues 206-224 following tryptic digestion of the hemagglutinin antigen. Specific peptide-antibody complexes formed after treatment of gel-recovered antigen are shown to be able to be preserved on the MALDI target array as has been previously demonstrated in this laboratory for whole virus. The approach has broad applicability for the analysis of a wide array of protein complexes with identification of the interaction interface in a single step with high-sample throughput and at low sample levels.
本文描述了一种基于蛋白质组学的方法，该方法结合了凝胶电泳和质谱，以高样本通量和灵敏度鉴定蛋白质相互作用及相互作用界面的性质。流感病毒蛋白质抗原的研究结果表明，该方法可成功用于检测当前流行的两种不同A型流感病毒血凝素抗原中的决定簇。这些决定簇在血凝素抗原经胰蛋白酶消化后定位于206-224位残基。如本实验室先前对完整病毒所证明的那样，凝胶回收抗原处理后形成的特异性肽-抗体复合物能够保存在MALDI靶阵列上。该方法对于分析各种蛋白质复合物具有广泛的适用性，能够在单一步骤中以高样本通量和低样本量鉴定相互作用界面。
