Ubiquitin at Fox Chase
Year: 2005
Type: article
Abstract: My interest in protein break-down as a research problem began in 1955 at about the time I joined the Biochemistry Department of Yale University. It was known that proteins break down intracellularly in the mature animal (1, 2). In 1955, I learned from Melvin Simpson about experiments he had published 2 years earlier (3) showing that a variety of conditions that should lower the ATP level of liver slices (anaerobiosis, cyanide, and 2,4-dinitrophenol) decreased the rate of liberation of labeled methionine from protein of rat liver slices. Simpson and I had just joined the Biochemistry Department, and we had labs down the hall from each other. Simpson's research goal at Yale was to look for an in vitro system that performed protein synthesis rather than protein breakdown. During my Yale years, I concentrated on enzyme mechanism questions relating to the keto-aldose isomerases, using tritium to establish the occurrence of proton transfer, but I kept an eye out for reports in the literature on progress in the protein breakdown field.
Nothing new was being reported by 1963, when we relocated to the Institute for Cancer Research at Fox Chase, Philadelphia. At that time, I began looking for a cell-free system that would show an ATP dependence using the Ehrlich ascites cells that were available from our animal colony. My usual procedure was to label cells in suspension with an essential amino acid, wash the cells, lyse them by homogenization, and look for acid-soluble counts by adding either ATP or 2-deoxyglucose plus hexokinase to deplete the endogenous ATP.
During 1972, on a half-year sabbatical divided between Oxford (J. Knowles) and Jerusalem (Y. Stein), I was able to do further experiments with tissues supplied by Hans Krebs and by Jacob Mager, who had been Avram Hershko's Ph.D. research director at the Hadassah …
Author: Irwin A. Rose
Institution: University of California, Irvine
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W1995202242