Ribosome Analysis In Vivo by Differential Scanning Calorimetry: The Effects of High Pressure on Escherichia coli

Abstract

We are developing differential scanning calorimetry (DSC) as a technique for ribosome analysis in vivo. This enables information to be obtained about ribosome conformation without cell disruption and using minimum cell processing. Using this method, we observed a linear correlation between cell death and the decrease in ribosome-associated enthalpy in pressure treated cells. This suggested a direct relationship between cell death and ribosome damage. Initially, the surviving ribosomes in pressure treated cells adopted a less stable conformation. During subsequent incubation at 37 °C, they restabilized over a period of several hours. The presence of elevated levels of extracellular Mg²⁺ during pressure treatment reduced ribosome destabilization.