title: Will heat-stable proteins nonspecifically protect cells from thermal stress?
abstract: In cell-free systems, stress-resistant proteins nonspecifically stabilize stress-susceptible proteins. This mechanism has been suggested to contribute to thermotolerance in cells (Minton et al.: Proc. Natl. Acad. Sci. USA, 79: 7107-7117, 1982). To test this hypothesis, red-blood-cell-mediated microinjection was used to transfer macromolecules into monolayers of CHO cells. We introduced the heat-stable proteins fetuin and ovomucoid into RBCs during hypotonic hemolysis and then fused the RBCs to CHO cells with polyethylene glycol as fusogen. Fetuin and ovomucoid were successfully transferred into 36-55% of the CHO cells as demonstrated by fluorescence of FITC-conjugated proteins. The plating efficiency of these CHO cells after fusion ranged from 35% to 60%. Three hours after fusion, CHO cells microinjected with fetuin or ovomucoid were exposed to 43 degrees C for 0-180 min or 45 degrees C for 0-40 min, and thermal survival was determined. There was no difference in cell survival between control untreated cells, control cells fused with nonloaded RBCs, and cells fused with RBCs loaded with fetuin or ovomucoid. While our results do not support the hypothesis that heat-stable proteins nonspecifically protect cells from thermal stress, several possible explanations are provided for this observation.
authors: Department of Radiation Oncology, University of California, San Francisco 94143.
authors: S W
authors: SW
authors: Prenninger
authors: G C
authors: GC
authors: Li
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival; D004912:Erythrocytes / Q000502:physiology*; D006360:Heat-Shock Proteins / Q000502:physiology*; D006358:Hot Temperature; D006801:Humans; D007700:Kinetics; D008845:Microinjections; D010059:Ovomucin / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D000509:alpha-Fetoproteins / Q000494:pharmacology
chemical_list: D006360:Heat-Shock Proteins; D000509:alpha-Fetoproteins; D012710:Serum Albumin, Bovine; D010059:Ovomucin