Abstract
Prion protein (PrP) is a ubiquitous conserved glycoprotein predominantly expressed in neurons of the central nervous system (CNS). To elucidate on its cellular function, we performed a yeast two-hybrid screen within an adult human brain cDNA library for potential PrP-binding molecules. A novel protein, HS-1 associated protein X-1 (HAX-1), was identified to be able to bind with PrP strongly. The interaction between the two proteins has been further verified by glutathione-S-transferase (GST) pull-down and immunoprecipitation assays. The minimal binding regions were mapped to the segments of residues aa 91–163 for PrPC and residues aa 38–129 for HAX-1. Immunofluorescent assays of co-expressions of human PrP and HAX-1 in 293T and SHSY-5Y cells revealed marked co-localizations of those two proteins in cytoplasm. Moreover, the co-expression of HAX-1 and wild-type PrP (PG5) was found to enhance the cellular resistance to the challenge of H2O2. Contrarily, co-transfection of HAX-1 did not reverse but aggravated the cytotoxicities of the genetic CJD (gCJD) associated PrP mutants with nine- (PG9) and fourteen-octarepeats (PG14). Our data provide for the first time a new PrP-interacting partner that may play role in cell oxidative stress and anti-apoptosis physiologically and cell damage pathologically.
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Acknowledgements
This work was supported by Chinese National Natural Science Foundation Grants 30771914 and 30800975, Institution Technique R&D Grant (2008EG150300), National Basic Research Program of China (973 Program) (2007CB310505), China Mega-Project for Infectious Disease (2009ZX10004-101), and the SKLID Development Grant (2008SKLID102 and 2008SKLID202).
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Supplemental Table 1
The primers for amplifying various lengths of human PrP and HAX-1 (DOC 45 kb)
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**g, YY., Li, XL., Shi, Q. et al. A Novel PrP Partner HS-1 Associated Protein X-1 (HAX-1) Protected the Cultured Cells Against the Challenge of H2O2 . J Mol Neurosci 45, 216–228 (2011). https://doi.org/10.1007/s12031-011-9498-2
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DOI: https://doi.org/10.1007/s12031-011-9498-2