1. 蛋白質體學：
利用蛋白質體微陣列晶片進行蛋白質體相關研究，特別是蛋白質體的交互作用。本實驗室除了具有大腸桿菌 (~4000 proteins) 蛋白質體微陣列晶片外，也具有酵母菌 (~5000 proteins) 和人類蛋白質體 (~18000 proteins) 微陣列晶片。
Our lab has a new high-throughput protein purification technique to purify ~4000 E. coli K12 proteins within ten hours. The purified proteins were then spotted on glass slides to form an E. coli K12 proteome chip. It was successfully applied to many projects. First, new DNA damage recognition activities in Escherichia coli were identified. It was also showed an important DNA repair protein; ybaZ (ATL) binds abasic DNA strongly and this result was published in Nature Methods. In addition to DNA repair study, we also set out to perform human serum profiling for the identification of biomarkers in inflammatory bowel diseases and for characterizing the ontogeny of anti-E.coli antibodies using human sera derived from infants and adults. Part of the result was published in Molecular and Cellular Proteomics.
利用大腸桿菌蛋白質體微陣列晶片探討抗菌肽於細菌內部的作用機制，也用於探討大腸桿菌蛋白質體和人體的互動關係 (Host & Microbe Interaction)。
We attempted to find the intracellular target of natural antimicrobial peptides by our E. coli K12 proteome chips. Until now, we already discovered several intracellular proteins and verified them to ensure their importance for antimicrobial to inhibit cell growth. We also use bioinformatics analysis to understand the intracellular activity of antimicrobial peptides. Particularly, we elucidated the relationships between two response regulators belonging to two-component system (TCS) with Lactoferricin B. TCS is highly conserved among all the microorganisms and control the gene expressions for bacteria to grow under their unique niche environments. Our study showed that Lfcin B binds to two response regulators, BasR and CreB, of TCS. For further analysis, we conducted several in vitro and in vivo experiments and utilized bioinformatics methods. The electrophoretic mobility shift assays and kinase assays indicate that Lfcin B inhibits the phosphorylation of the response regulators (BasR and CreB) and their cognate sensor kinases (BasS and CreC). Antibacterial assays showed that Lfcin B reduced E. coli’s tolerance to environmental stimuli, such as excessive ferric ions and minimal medium conditions. This is the first study to show that an antimicrobial peptide attacks TCS directly. This study is in press by Molecular and Cellular Proteomics.
In addition to E. coli proteome chip, our lab also has yeast and human proteome microarrays. Thus, we are expending our research area from E. coli to yeast and human as well.
2. 奈米生醫感測科技
應用奈米生化科技在多樣免疫分析技術 (simultaneous multiplexed immunoassays) 也是我們研究的重點。我們專長以蛋白質G-奈米脂粒 (protein G-liposomal nanovesicles) 以作為免疫分析的擴大訊號通用標劑 (universal reagent)。利用奈米脂粒，我們發展的感測科技平台為高通量的盤式系統和微陣列系統。所檢測的分子從小分子的抗生素到大分子的疼痛分子，甚至是細菌都是我們奈米生醫感測科技的檢測興趣範圍。
