Conformational change of glutathione-S-transferase by its co-expression with prion domain of yeast Ure2p

【摘要】 <正> The Ure2 protein from Saccharomyces cerevisisae has a changeable structure similar to that of "mammalian prion protein. Its N-terminal is the prion domain (PrD) consisting of 65 amino acids which plays a critical role in yeast prion development. In this study, PrD gene was recombinated with glutathione-S-transferase(GST) gene, and a soluble GST-PrD(sGST-PrD) fusion protein was expressed in E. -coli. sGST-PrD could spontaneously polymerize into amyloid fibrils in vitro, displaying typical ^-sheet-type structure; it had increased resistance to proteinase K and exhibited amyloid-like optical properties. Moreover, the aggregated GST-PrD(aGST-PrD) could induce sGST-PrD to aggregate into fibrils. These results indicate that PrD could change the conformation of GST moiety in a recombinant protein with PrD to form a prion-like chimeric protein, which proves that PrD has the ability to mediate a prion-like conversion of other proteins fused with it.更多还原

【Abstract】 The Ure2 protein from Saccharomyces cerevisisae has a changeable structure similar to that of "mammalian prion protein. Its N-terminal is the prion domain (PrD) consisting of 65 amino acids which plays a critical role in yeast prion development. In this study, PrD gene was recombinated with glutathione-S-transferase(GST) gene, and a soluble GST-PrD(sGST-PrD) fusion protein was expressed in E. -coli. sGST-PrD could spontaneously polymerize into amyloid fibrils in vitro, displaying typical ^-sheet-type structure; it had increased resistance to proteinase K and exhibited amyloid-like optical properties. Moreover, the aggregated GST-PrD(aGST-PrD) could induce sGST-PrD to aggregate into fibrils. These results indicate that PrD could change the conformation of GST moiety in a recombinant protein with PrD to form a prion-like chimeric protein, which proves that PrD has the ability to mediate a prion-like conversion of other proteins fused with it.更多还原