effect of hydrophobic mutations in the h2-h3 subdomain of prion protein on stability and conversion in vitro and in vivo疏水效应h2-h3突变子域名朊蛋白的稳定性和转换在体外和体内.pdfVIP

Effect of Hydrophobic Mutations in the H2-H3 Subdomain of Prion Protein on Stability and Conversion In Vitro and In Vivo ˇ 1. 2. 1.¤ ˇ 3 4 Iva Hafner-Bratkovic , Lars Gaedtke , Andrej Ondracka , Peter Veranic , Ina Vorberg , Roman Jerala1,5,6* ¨ 1 Department of Biotechnology, National Institute of Chemistry, Ljubljana, Slovenia, 2 Institute of Virology, Technical University Munich, Munchen, Germany, 3 Institute of Cell Biology, School of Medicine, University of Ljubljana, Ljubljana, Slovenia, 4 German Centre for Neurodegenerative Diseases (DZNE), Bonn, Germany, 5 Faculty of Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, Slovenia, 6 ENR FIST Centre of Excellence, Ljubljana, Slovenia Abstract Prion diseases are fatal neurodegenerative diseases, which can be acquired, sporadic or genetic, the latter being linked to mutations in the gene encoding prion protein. We have recently described the importance of subdomain separation in the conversion of prion protein (PrP). The goal of the present study was to investigate the effect of increasing the hydrophobic interactions within the H2-H3 subdomain on PrP conversion. Three hydrophobic mutations were introduced into PrP. The mutation V209I associated with human prion disease did not alter protein stability or in vitro fibrillization propensity of PrP. The designed mutations V175I and T187I on the other hand increased protein thermal stability. V175I mutant fibrillized faster than wild-type PrP. Conversion delay of T187I was slightly longer, but fluore