| Summary: | 碩士 === 國立陽明大學 === 生命科學暨基因體科學研究所 === 95 === Protein engineering has become a valuable tool for creating new proteins for practical use. In this study, we tried to design a preliminary protein drug, CPP-sarcin, which was expected to cross the plasma membrane and hydrolyze its target, the ribosome. α sarcin is a ribotoxin that inactivates the ribosome by cleaving 28S rRNA in eukaryotes. α sarcin itself is unable to enter the cell, and therefore we fused a cell penetrating peptide (CPP) with α sarcin. CPPs are a variety of cationic short peptides, which have an ability to make cell entry. In this study, three types of CPPs, namely Tat, Antennapedia, and Polyarginine, were experimentally fused to α sarcin. These CPP-sarcins were analyzed for their ribosome hydrolytic activity and cell entry activity. The result showed that TAT-sarcin was the best among the three. It was found that HeLa cells pretreated with TAT-sarcin for an hour showed in protein synthesis inhibition, cell morphology changes, and eventually cell death.
|
|---|
