NR AXLB
AU Dong,X.P.; Li,P.; Han,J.; Lei,Y.J.; Shan,B.; Dong,C.F.; Xiao,X.L.; Jiang,H.Y.; Gao,C.
TI Doppel Protein Induced Cytotoxic Effect on Neuron Cells in vitro and PrP Protein was Able to Antagonize Doppel-associated Cytotoxicity
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.110
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: Doppel (Dpl) is recently identified as a prion (PrP)-like protein due to the structural and biochemical similarities, however, the natural functions of Dpl and PrP remain unclear yet.
Objective: To assess the potential activity of proteins Dpl and PrP on neural cells in vitro.
Methods: A 531-bp human PRND gene sequence encoding Dpl protein was amplified from human peripheral blood leucocytes. The full-length and various truncated Dpl and PrP proteins were expressed and purified from Escherichia coli. Various proteins were supplied on the cultured cell line SH-SY5Y. The cell growth situations were evaluated by MTT and Traplan blue assays and apoptosis phenomenon were observed by staining of Hoechest33324, cytometry after staining of Annexin V/PI and TUNER assay.
Results: Supplement of the full-length Dpl protein onto the cultured human neuroblastoma cell line SH-SY5Y induced remarkable cytotoxicity and the region responsible for its cytotoxicity was mapped at the middle segment of Dpl (amino acid 81-122). Interestingly, employment of full-length wide type human PrP protein onto the cultured cells antagonized Dpl-induced cytotoxicity. Analysis on fragments of PrP mutants showed that its N-terminal fragment (amino acid 23-90) was responsible for the protective activity. Treatment of a truncated PrP (the full-length PrP from aa 23 to 231 in which the peptide from aa 32 to 231 is deleted), which shared similar secondary structure as Dpl, induced cytotoxicity on the cells like Dpl. Furthermore, copper ion could enhance the antagonizing effect of PrP on Dpl-induced cytotoxicity. Apoptosis assays revealed that cytotoxicity induced in Dpl occurred via an apoptotic mechanism.
Conclusion: These results suggested that function of Dpl is antagonistic to PrP rather than synergistic and are consistent with early observations in wide transgenic mice.
AD X.-P. Dong, P. Li, J. Han, Y.-J. Lei, B. Shan, C.-F. Dong, X.-L. Xiao, H.-Y. Jiang, C. Gao, China CDC, State Key Laboratory for Infectious Disease Preven, China
SP englisch
PO Schottland