Denaturation of proteins during heat shock. In vivo recovery of solubility and activity of reporter enzymes.
Using beta-galactosidase and luciferase as reporter enzymes, we have previously shown that enzymatic inactivation occurring during a heat shock is concomitant with protein insolubilization (Nguyen, V. T., Morange, M., and Bensaude, O. (1989) J. Biol. Chem. 264, 10487-10492). In this paper, we observe that pretreatment of cells with D2O and glycerol, compounds known to stabilize protein structure, leads to a parallel decrease of protein inactivation and insolubilization, suggesting that these two phenomena result most probably from heat-induced protein denaturation. We found that heat shock-promoted inactivation and insolubilization are not irreversible processes, since even in the absence of protein synthesis, beta-galactosidase solubility and luciferase solubility and activity are recovered in vivo after a heat treatment. Cognate heat shock proteins might be involved in this renaturation process.
Pinto
M
M
Groupe de Biologie Moléculaire du Stress, Institut Pasteur, Paris, France.
Morange
M
M
Bensaude
O
O
D000818:Animals; D002478:Cells, Cultured; D005990:Glycerol / Q000737:chemistry; D006358:Hot Temperature; D066298:In Vitro Techniques; D007739:L Cells; D008156:Luciferases / Q000737:chemistry*; D051379:Mice; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000737:chemistry; D012995:Solubility; D001616:beta-Galactosidase / Q000737:chemistry*
D011994:Recombinant Proteins; D008156:Luciferases; D001616:beta-Galactosidase; D005990:Glycerol