Molecular interaction between prion protein and GFAP both in native and recombinant forms in vitro.
Gliosis of glial fibrillary acidic protein (GFAP) associated astrocytes is considered to be one of the hallmarks of transmissible spongiform encephalopathies (TSEs). In the present study, remarkable GFAP-PrP(Sc) or GFAP-PrP(C) complexes were separately detected in the brain homogenates of 263 K (Scrapie)-infected or normal hamsters by co-immunoprecipitation assay. To get more exact molecular evidences for interaction between prion protein (PrP) and GFAP, various recombinant PrP or GFAP proteins were expressed using prokaryotic-expressing and in vitro translation system. Using pull down and co-immunoprecipitation assays, reliable molecular interaction between PrP and GFAP was observed, and proteinase K (PK)-digested PrP(Sc) molecules were confirmed to be able to bind the recombinant GFAP specifically as well. The region within PrP that was responsible for interaction with GFAP was narrowed to PK-resistant core of PrP (i.e. aa 91-230). The study of the association of PrP with GFAP supplies the molecular evidence for the observation of co-localization of PrP(Sc) and GFAP in the brains of TSEs and may further provide insight into a potential role of GFAP in the biological function of PrP and the pathogenesis of prion diseases.
Dong
Chen-Fang
CF
State Key Laboratory for Infectious Disease Prevention and Control, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Ying-Xin Rd 100, 100052, Beijing, People's Republic of China.
Wang
Xiao-Fan
XF
Wang
Xin
X
Shi
Song
S
Wang
Gui-Rong
GR
Shan
Bing
B
An
Run
R
Li
Xiao-Li
XL
Zhang
Bao-Yun
BY
Han
Jun
J
Dong
Xiao-Ping
XP
D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism / Q000473:pathology; D006224:Cricetinae; D019286:Endopeptidase K / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism*; D047468:Immunoprecipitation; D051379:Mice; D018621:PrPC Proteins / Q000235:genetics / Q000378:metabolism*; D018620:PrPSc Proteins / Q000235:genetics / Q000378:metabolism*; D017096:Prion Diseases / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*
D005904:Glial Fibrillary Acidic Protein; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011994:Recombinant Proteins; D019286:Endopeptidase K