Permeabilization of yeast cells: Application to study on the regulation of AMP deaminase activity in situ

Abstract A permeabilization method which allows the assay of several intracellular enzymes within the boundaries of the yeast cell wall is described. Toluene treatment was found to make yeast cells completely permeable to exogenous substrates, and intracellular enzymes did not leak out of the treated cells. This method was also compared with the permeabilization techniques reported previously. Electron microscopic examination of toluene-treated cells indicated that they were essentially intact. The kinetic properties of AMP deaminase, examined in the permeabilized cells, including allosteric regulation by polyamine and Zn 2+ , suggest some differences in protein interactions for AMP deaminase in situ and in vitro .