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PLoS One. 2012;7(11):e49021. doi: 10.1371/journal.pone.0049021. Epub 2012 Nov 8.

Identification of proteins sensitive to thermal stress in human neuroblastoma and glioma cell lines.

Author information

1
Department of Neuroscience, Santa Fe Health Alzheimer's Disease Research Center, University of Florida, Gainesville, FL, USA. xugl@ufl.edu

Erratum in

  • PLoS One. 2012;7(11). doi:10.1371/annotation/82b96c01-6435-4856-80a6-0176b1986e32. Kobiessy, Firas [corrected to Kobeissy, Firas].

Abstract

Heat-shock is an acute insult to the mammalian proteome. The sudden elevation in temperature has far-reaching effects on protein metabolism, leads to a rapid inhibition of most protein synthesis, and the induction of protein chaperones. Using heat-shock in cells of neuronal (SH-SY5Y) and glial (CCF-STTG1) lineage, in conjunction with detergent extraction and sedimentation followed by LC-MS/MS proteomic approaches, we sought to identify human proteins that lose solubility upon heat-shock. The two cell lines showed largely overlapping profiles of proteins detected by LC-MS/MS. We identified 58 proteins in detergent insoluble fractions as losing solubility in after heat shock; 10 were common between the 2 cell lines. A subset of the proteins identified by LC-MS/MS was validated by immunoblotting of similarly prepared fractions. Ultimately, we were able to definitively identify 3 proteins as putatively metastable neural proteins; FEN1, CDK1, and TDP-43. We also determined that after heat-shock these cells accumulate insoluble polyubiquitin chains largely linked via lysine 48 (K-48) residues. Collectively, this study identifies human neural proteins that lose solubility upon heat-shock. These proteins may represent components of the human proteome that are vulnerable to misfolding in settings of proteostasis stress.

PMID:
23145051
PMCID:
PMC3493505
DOI:
10.1371/journal.pone.0049021
[Indexed for MEDLINE]
Free PMC Article

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