Bounhar Y, Zhang Y, Goodyer CG, LeBlanc A. Prion protein protects human neurons against Bax-mediated apoptosis. J Biol Chem. 2001 Oct 19;276(42):39145-9. PubMed Abstract

REAGENTS/MATERIAL:
Human Bcl-2 cNDA, kind gift from Dr. Walter Nishioka (Vical Inc. San Diego, CA) PrP and PrP-D178N and -T183A DNAs were PCR-amplified. Bax-a cDNA was PCR-amplified from human neuron cNDA. cDNAs were cloned into pBluescript KSII (pBSK-; Stratagene) or pCep4b (Invitrogen) Deletion of OR region by PCR amplification with the following primers: (PDG2) 5'-TACTGAGAATTCGCAGTCCATTATGGCGAACCTTGGCTGCTGG-3' (P3-) 5'-ACCACCGCCCTGAGG-3' (P4+) 5'-ACCCACAGTCAGTGG-3' (PDG1) 5'GTACTGAGGATCCTCCTCATCCCACTATCAGGAAGA-3' Human primary neurons were cultured and microinjected with 25 pl containing 0.75 pg of DNA and 2.5 pg of dextran Texas Red in phosphate buffered saline.

FUTURE DIRECTION:
Further studies needed to understand the underlying molecular mechanism of PrP function against Bax. Also, to determined whether FFI and GASE PrP mutation undergo a loss of function against Bax-mediated cell death is due to altered protein conformation or improper trafficking.