Classics: The Purification of Diphtheria Toxin by Alwin M. Pappenheimer, Jr. Robert D. Simoni, Robert L. Hill and Martha Vaughan J. Biol. Chem. 2004, 279:e4. Find articles, minireviews, Reflections and Classics on similar topics on the JBC Affinity Sites. Alerts: • When this article is cited • When a correction for this article is posted Click here to choose from all of JBC's e-mail alerts This article cites 5 references, 4 of which can be accessed free at http://www.jbc.org/content/279/4/e4.full.html#ref-list-1 Downloaded from http://www.jbc.org/ by guest on November 14, 2014 Access the most updated version of this article at http://www.jbc.org/content/279/4/e4 THE JOURNAL OF BIOLOGICAL CHEMISTRY © 2004 by The American Society for Biochemistry and Molecular Biology, Inc. Vol. 279, No. 4, Issue of January 23, p. e4, 2004 Printed in U.S.A. Classics A PAPER IN A SERIES REPRINTED TO CELEBRATE THE CENTENARY OF THE JBC IN 2005 JBC Centennial 1905–2005 100 Years of Biochemistry and Molecular Biology The Purification of Diphtheria Toxin by Alwin M. Pappenheimer, Jr. Diphtheria Toxin. I. Isolation and Characterization of a Toxic Protein from Cornynebacterium Diphtheriæ Filtrates (Pappenheimer, A. M., Jr. (1937) J. Biol. Chem. 120, 543–553) 1 All biographical material was taken from this source. This paper is available on line at http://www.jbc.org 95 Downloaded from http://www.jbc.org/ by guest on November 14, 2014 Alwin M. Pappenheimer Jr. (1908 –1995) was born in Cedarhurst, New York. His father, Alwin M. Pappenheimer, was a distinguished pathologist at the College of Physicians and Surgeons at Columbia University. Among his father’s many contributions was his discovery of the link between light and bone deposition, which was published in a previous JBC Classic (1). Pappenheimer grew up in an intellectual household and was interested in science from an early age. He entered Harvard in 1925 at the age of 17; he was among the first students to enroll in the new biochemical sciences tutorial. Appropriately, he later returned to Harvard and succeeded John T. Edsall, author of a previous JBC Classic (2), as Chairman of the Board of Tutors of the Biochemical Sciences Program. After choosing graduate school rather than medical school, Pappenheimer entered the graduate program in organic chemistry at Harvard with James B. Conant as his advisor even though he was determined already at the time to have a career in biological research. When he completed his Ph.D. work in 1932, in the midst of the Great Depression, although jobs were difficult to find he was able to spend 1 year as a postdoctoral fellow with Hans Zinsser at Harvard studying pneumococcal polysaccharides. This was followed by a 2-year National Research Council Fellowship with Sir Henry Dale at the National Institute of Medical Research in London working on the isolation of a bacterial growth factor called “sporogenes vitamin.” This experience sparked his future interests in isolation of a bacterial toxin. In 1935, Pappenheimer returned to Cambridge, Massachusetts without a job but with the plan in mind “to isolate a pure potent bacterial toxin and to find out what makes it so toxic” (3).1 He discussed his ideas with J. Howard Mueller in the Department of Bacteriology at Harvard, another author of a previous JBC Classic (4). Mueller helped him to obtain a Bradford Fellowship at Harvard. He was provided space and some technical support at the Jamaica Plains antitoxin and vaccine laboratory where the work reported in this JBC Classic was done. Attempts to purify diphtheria toxin had been frustrated because the bacterial culture medium from which the toxin was to be isolated contained a complex mixture of protein, and the purification of toxin free of contaminating culture components had been proven impossible. Pappenheimer set out to devise a better defined culture medium without high molecular weight ingredients to interfere with toxin purification. He was successful in optimizing growth conditions and succeeded in the complete purification of the toxin, as reported in this JBC Classic. The modification of the growth medium had ensured that it contained no ammonium sulfate-precipitable material permitting separation and purification of the toxin by differential ammonium sulfate fractionation. This was the first bacterial toxin purified in crystalline form and brought Pappenheimer great international recognition. In 1939, Pappenheimer moved to the University of Pennsylvania as an Assistant Professor of Bacteriology but after only 2 years was recruited to the Department of Bacteriology at New 96 Classics York University (NYU) by the new chairman Colin E. MacLeod. MacLeod and Pappenheimer built a new Department of Microbiology while the latter continued his work on bacterial growth and toxin production. His research was interrupted by his service as an Army captain in World War II but resumed when he returned to NYU in 1945. After MacLeod left NYU for the University of Pennsylvania, Pappenheimer became chairman of the Department of Microbiology, but John T. Edsall soon recruited him to return to Harvard as Professor of Biology and Chairman of the Board of Tutors in Biochemical Sciences, the program in which he had been enrolled as an undergraduate. After achieving his first goal with the purification of diphtheria toxin, Pappenheimer, his students, and others worked on discovering the mechanism of its toxicity. With R. John Collier, he showed that diphtheria toxin inhibited protein synthesis in HeLa cells and in HeLa cell extracts (5). Then Collier in 1966 –1967 showed that elongation factor 2, EF-2, was inactivated by diphtheria toxin in the presence of NAD and in 1968 –1969 D. Michael Gill described the ADP-ribosylation reaction catalyzed by diphtheria toxin, which results in the inactivation of EF-2. Pappenheimer received many honors for his work including the Eli Lilly Award in 1942 and election to the National Academy of Sciences in 1973. Together with his former student R. John Collier, Pappenheimer was awarded the Paul Ehrlich Prize and Gold Medal. Robert D. Simoni, Robert L. Hill, and Martha Vaughan REFERENCES 1. JBC Classics: Hess, A. F., Unger, L. J., and Pappenheimer, A. M. (1922) J. Biol. Chem. 50, 77– 81 (http://www.jbc.org/cgi/content/full/277/23/e12) 2. JBC Classics: Edsall, J. T. (1930) J. Biol. Chem. 89, 289 –313; Cohn, E. J., Edsall, J. T., and Blanchard, M. H. (1933) J. Biol. Chem. 105, 319 –326 (http://www.jbc.org/cgi/content/full/277/33/29351) 3. Lawrence, H. S. (1999) Biographical memoir of Alwin Max Pappenheimer, Jr. Natl. Acad. Sci. 77, 3–18 4. JBC Classics: Mueller, J. H. (1923) J. Biol. Chem. 56, 157–169 (http://www.jbc.org/cgi/content/full/277/25/e14) 5. Collier, R. J., and Pappenheimer, A. M. (1964) J. Exp. Med. 120, 1007–1018, 1019 –1039 Downloaded from http://www.jbc.org/ by guest on November 14, 2014 Alwin M. Pappenheimer, Jr. Photo courtesy of the National Library of Medicine.
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