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The Rockefeller Institute and Albert Claude
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The Rockefeller Institute and Albert Claude
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Views | Duration | ||
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21. Writing to Carl Cori, and crystallised insulin | 202 | 03:14 | |
22. The Lilly Research Labs: Making a re-discovery | 202 | 03:40 | |
23. 'You will get a fur coat when I get a Nobel Prize' | 229 | 05:01 | |
24. Becoming Professor of Biochemistry at Louvain | 185 | 01:52 | |
25. Setting up a lab: A grant from Eli Lilly and Company | 151 | 02:40 | |
26. Setting up a lab: Difficulties getting equipment | 148 | 02:51 | |
27. Glucose 1-phosphate and glucose 6-phosphate | 184 | 03:55 | |
28. Purifying enzymes and visiting labs | 144 | 03:05 | |
29. The Rockefeller Institute and Albert Claude | 1 | 206 | 04:34 |
30. Centrifugal fractionation | 187 | 02:12 |
Then we had to purify the enzyme. Well, in those days, there were not so many techniques for purifying protein enzymes and one of the simplest techniques was what is known as isoelectric precipitation, which takes advantage of the fact that proteins are least soluble when their electric charge is zero. And so you acidify the medium and you reach a pH where many proteins will precipitate because of this lack of electric charge. And so at pH five we got a big precipitate and our enzyme was there in the precipitate. So that was the first step: a very dirty precipitate, obviously. And so the next step was, of course... if this was isoelectric precipitation, was to re-dissolve the enzyme and try something else, like ammonium sulphate precipitation or alcohol precipitation which were the only techniques available at that time. Chromatography was not known yet, or at least not for proteins. And so we took our precipitate, changed the pH back to seven; enzyme would not re-dissolve. So this was not the reversible kind of reaction that you have with iso-electro precipitation; this was an irreversible agglutination, which implied that the enzyme was probably associated with some kind of cell structure. It so happens that on my return from the United States... some time before that, I had visited a few labs and I had visited the best: the discoverer of insulin in Toronto; I had visited a man called Levine in Chicago because he'd worked on insulin and liver; I'd visited Lipmann in Boston because he was one of my heroes – Lipmann the father of bioenergetics. I was so impressed with this work, and I found it so beautiful I went to see him. You know, amazingly, I wouldn't do this anymore... today; a young student would come to me and say, 'I want to see you because I admire you'; I would say, 'Well I have more interesting or important things to do.' No, he actually... he saw me, talked with me, even invited me and my wife to a Chinese restaurant. So I had a lovely time in Boston – finally ended up in New York where I went to the Rockefeller Institute for Medical Research.
Belgian biochemist Christian de Duve (1917-2013) was best known for his work on understanding and categorising subcellular organelles. He won the Nobel Prize in Physiology or Medicine in 1974 for his joint discovery of lysosomes, the subcellular organelles that digest macromolecules and deal with ingested bacteria.
Title: Purifying enzymes and visiting labs
Listeners: Peter Newmark
Peter Newmark has recently retired as Editorial Director of BioMed Central Ltd, the Open Access journal publisher. He obtained a D. Phil. from Oxford University and was originally a research biochemist at St Bartholomew's Hospital Medical School in London, but left research to become Biology Editor and then Deputy Editor of the journal Nature. He then became Managing Director of Current Biology Ltd, where he started a series of Current Opinion journals, and was founding Editor of the journal Current Biology. Subsequently he was Editorial Director for Elsevier Science London, before joining BioMed Central Ltd.
Tags: Fritz Lipmann, Rachmiel Levine
Duration: 3 minutes, 5 seconds
Date story recorded: September 2005
Date story went live: 24 January 2008