Interaction of Doppel with the full-length laminin receptor precursor protein.
Doppel (Dpl) is a homolog of normal cellular prion protein (PrPc) with unknown functions. Ectopic expression of Dpl in the central nervous system (CNS) causes neurotoxicity and this effect is rescued by the expression of PrPc. However, the molecular basis for the protective effect of PrPc remains unclear. Using a yeast two-hybrid system, we showed that Dpl binds the full-length 37-kDa laminin receptor precursor protein (LRP), one of the receptors of PrPc. The interaction was also validated by immunoprecipitation and immunoblotting using transfected cell lines and in vivo derived tissues. Further mapping experiments showed that although the middle fragment containing residues 100-220 of LRP was able to interact with Dpl, deletion of the N-terminal domain of the full-length LRP abolished its interaction with Dpl. These results suggest that while both PrPc and Dpl interact with LRP, the domains that are involved in the binding are not the same. Our results may have implications for the molecular mechanisms of Dpl-PrPc antagonism and physiological roles of Dpl.
Yin
Shao-Man
SM
Institute of Microbiology, Chinese Academy of Science, Beijing 100080, People's Republic of China.
Sy
Man-Sun
MS
Yang
Huai-Yi
HY
Tien
Po
P
D000818:Animals; D002460:Cell Line; D002490:Central Nervous System / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D005260:Female; D058851:GPI-Linked Proteins; D017353:Gene Deletion; D006801:Humans; D015151:Immunoblotting; D008024:Ligands; D008297:Male; D051379:Mice; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D011233:Precipitin Tests; D011328:Prions / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D017474:Receptors, Laminin / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D013737:Testis / Q000378:metabolism / Q000473:pathology; D014162:Transfection; D020798:Two-Hybrid System Techniques
D018076:DNA, Complementary; D058851:GPI-Linked Proteins; D008024:Ligands; C501125:PRND protein, human; D011328:Prions; C122036:Prnd protein, mouse; D011498:Protein Precursors; D017474:Receptors, Laminin; D011994:Recombinant Proteins