Permeabilization of yeast cells: Application to study on the regulation of AMP deaminase activity in situ |
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Authors: | Keiko Murakami Hiroshi Nagura Masataka Yoshino |
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Institution: | 1. Department of Biochemistry, Yokohama City University School of Medicine, Yokohama 232 Japan;2. Department of Pathology, Tokai University School of Medicine, Isehara, Kanagawa 259-11, Japan |
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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 Zn2+, suggest some differences in protein interactions for AMP deaminase in situ and in vitro. |
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Keywords: | To whom correspondence should be addressed |
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