Effects of recombinant protein misfolding and aggregation on bacterial membranes.
The expression of recombinant proteins is known to induce a metabolic rearrangement in the host cell. We used aggregation-sensitive model systems to study the effects elicited in Escherichia coli cells by the aggregation of recombinant glutathione-S-transferase and its fusion with the green fluorescent protein that, according to the expression conditions, accumulate intracellularly as soluble protein, or soluble and insoluble aggregates. We show that the folding state of the recombinant protein and the complexity of the intracellular aggregates critically affect the cell response. Specifically, protein misfolding and aggregation induce changes in specific host proteins involved in lipid metabolism and oxidative stress, a reduction in the membrane permeability, as well as a rearrangement of its lipid composition. The temporal evolution of the host cell response and that of the aggregation process pointed out that the misfolded protein and soluble aggregates are responsible for the membrane modifications and the changes in the host protein levels. Interestingly, native recombinant protein and large insoluble aggregates do not seem to activate stress markers and membrane rearrangements.
Ami
D
D
Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, 20126 Milan, Italy.
Natalello
A
A
Schultz
T
T
Gatti-Lafranconi
P
P
Lotti
M
M
Doglia
S M
SM
de Marco
A
A
D002462:Cell Membrane / Q000502:physiology*; D002463:Cell Membrane Permeability; D004926:Escherichia coli / Q000201:enzymology*; D017930:Genes, Reporter; D005982:Glutathione Transferase / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008563:Membrane Lipids / Q000378:metabolism; D018384:Oxidative Stress; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D017550:Spectroscopy, Fourier Transform Infrared; D001616:beta-Galactosidase / Q000235:genetics
D008563:Membrane Lipids; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005982:Glutathione Transferase; D001616:beta-Galactosidase