title: Protein denaturation during heat shock and related stress. Escherichia coli beta-galactosidase and Photinus pyralis luciferase inactivation in mouse cells.
abstract: In an attempt to question the toxic effect of heat shock and related stress, we have studied the activity of reporter enzymes during stress. Escherichia coli beta-galactosidase and Photinus pyralis luciferase were synthesized in mouse and Drosophila cells after transfection of the corresponding genes. Both enzymes are rapidly inactivated during hyperthermia. The corresponding polypeptides are not degraded but become insoluble even in the presence of non-ionic detergents. The heat inactivation is more dramatic in vivo within the living cell than in vitro, in a detergent-free crude cell lysate. The extent of enzyme inactivation at a given temperature depends on the cell type in which the enzyme is expressed. Luciferase is inactivated at lower temperatures within Drosophila cells than within mouse cells, whereas beta-galactosidase is inactivated at higher temperatures in E. coli than in mouse cells. A "priming" heat shock confers a transient increased resistance (thermotolerance) of cells against a second "challenging" heat shock. Enzyme inactivation during heat shock or exposure of the cells to ethanol is attenuated in heat shock-primed cells. A comparable thermoprotection is raised by a priming heat shock for both luciferase activity and protein synthesis. Thus, the study of reporter enzyme inactivation is a promising tool for understanding the molecular basis of the toxicity of heat shock and related stress as well as the mechanisms leading to thermotolerance.
authors.affiliation: Groupe de Biologie Moléculaire du Stress, Institut Pasteur, Paris, France.
authors.forename: V T
authors.initials: VT
authors.lastname: Nguyen
authors.forename: M
authors.initials: M
authors.lastname: Morange
authors.forename: O
authors.initials: O
authors.lastname: Bensaude
mesh_terms: D000818:Animals; D001517:Coleoptera / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005696:Galactosidases / Q000037:antagonists & inhibitors*; D005796:Genes; D005798:Genes, Bacterial; D006358:Hot Temperature; D007700:Kinetics; D008156:Luciferases / Q000037:antagonists & inhibitors*; D051379:Mice; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D013816:Thermodynamics; D014162:Transfection; D001616:beta-Galactosidase / Q000037:antagonists & inhibitors*
chemical_list: D011994:Recombinant Proteins; D008156:Luciferases; D005696:Galactosidases; D001616:beta-Galactosidase