RRC ID 72160
Author Satoh J, Onoue H, Arima K, Yamamura T.
Title The 14-3-3 protein forms a molecular complex with heat shock protein Hsp60 and cellular prion protein.
Journal J Neuropathol Exp Neurol
Abstract The 14-3-3 protein family consists of acidic 30-kDa proteins composed of 7 isoforms expressed abundantly in neurons and glial cells of the central nervous system (CNS). The 14-3-3 protein identified in the cerebrospinal fluid provides a surrogate marker for premortem diagnosis of Creutzfeldt-Jakob disease, although an active involvement of 14-3-3 in the pathogenesis of prion diseases remains unknown. By protein overlay and mass spectrometric analysis of protein extract of NTera2-derived differentiated neurons, we identified heat shock protein Hsp60 as a 14-3-3-interacting protein. The 14-3-3zeta and gamma isoforms interacted with Hsp60, suggesting that the interaction is not isoform-specific. Furthermore, the interaction was identified in SK-N-SH neuroblastoma, U-373MG astrocytoma, and HeLa cervical carcinoma cells. The cellular prion protein (PrPC) along with Hsp60 was coimmunoprecipitated with 14-3-3 in the human brain protein extract. By protein overlay, 14-3-3 interacted with both recombinant human Hsp60 and PrPC produced by Escherichia coli, indicating that the molecular interaction is phosphorylation-independent. The 14-3-3-binding domain was located in the N-terminal half (NTF) of Hsp60 spanning amino acid residues 27-287 and the NTF of PrPC spanning amino acid residues 23-137. By immunostaining, the 14-3-3 protein Hsp60 and PrPC were colocalized chiefly in the mitochondria of human neuronal progenitor cells in culture, and were coexpressed most prominently in neurons and reactive astrocytes in the human brain. These observations indicate that the 14-3-3 protein forms a molecular complex with Hsp60 and PrPC in the human CNS under physiological conditions and suggest that this complex might become disintegrated in the pathologic process of prion diseases.
Volume 64(10)
Pages 858-68
Published 2005-10-1
DOI 10.1097/01.jnen.0000182979.56612.08
PII 00005072-200510000-00004
PMID 16215457
MeSH 14-3-3 Proteins / genetics 14-3-3 Proteins / metabolism* Amino Acid Sequence Brain / metabolism* Cell Line Chaperonin 60 / chemistry Chaperonin 60 / genetics Chaperonin 60 / metabolism* Electrophoresis, Gel, Two-Dimensional Humans Immunohistochemistry Immunoprecipitation Mass Spectrometry Molecular Sequence Data Neurons / metabolism* Peptide Fragments / metabolism Phosphorylation PrPC Proteins / metabolism* Protein Structure, Tertiary Recombinant Proteins / metabolism
IF 2.923
Human and Animal Cells 293(RCB1637) SK-N-SH(RCB0426)