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SIDNEY UDENFRIEND Abri;t 5, ~ 9 ~ 8-December 29, ~ 999 BY HERBERT WEISSBACH AND BERNHARD WITKOP SIDNEY UDENFRIEND S PARENTS emigrated to the United States from an Austro-Polish region in central Eu- rope in 1913. They had three children, the oldest was Sidney, who was born in Brooklyn, New York, on April 5, 1918. After attending public schools in Brooklyn Uclenfrienc! en- terecl the City College of New York (CCNY) in 1935. At that time CCNY was the ciream for so many of the immigrant parents who wantec! their chiTciren to obtain a college eclu- cation. Supported by public funcis, with no tuition, CCNY proviclecl that opportunity for those students who couIcl pass the rigid requirements for entrance. The Chemistry De- partment was well recognized in the field of physiological chemistry (or biochemistry) thanks in large part to Ben- jamin Harrow, who wrote a widely used textbook. Harrow hacl a great influence on UclenfriencI, en cl after graduation in 1939 Uclenfriencl was set on a career in bio- chemistry and determined to go to graduate school. In 1940 he was accepted at New York University Graduate School in the Department of Biology working with Kenneth BlancharcI. At nights he hac! a position with the New York City Depart- ment of Health directing other graduate students in carry- ing out Wasserman tests on draftees for the Army. In 1942 271
272 BIOGRAPHICAL MEMOIRS he received his M.S. degree and with the country at war, he took a position as a biochemist in the New York University malaria program at Goldwater Memorial Hospital in New York City. James Shannon directed this program, and Udenfriend was placed in a group headed by Bernard B. ("Steve") Brodie. His research involved developing new ana- Tytical methods for drugs and studying drug metabolism (1943~. The malaria program was considered vital defense research and Udenfriend was deferred from the draft, and obtained valuable research experience in this exciting envi- ronment until the end of the war. During this neriod at ~ . . . - - ~ - -- a Goldwater he married Shirley Reidel. They remained to- gether for 56 years, until his death in 1999, and they had two children, daughter Aliza and son Elliot. In the fall of 1945 he returned to New York University to complete his graduate studies, initially working with Severo Ochoa in the Department of Biochemistry of the Medical School. Ochoa left the department after one year, and Udenfriend changed mentors and continued his thesis work with Albert Keston. Together they developed the isotope- derivative method for the assay of amino acids and for de- termining amino terminal residues in proteins (1949~. He received his Ph.D. in biochemistry from New York Univer- sity in 1948 and accepted a position as instructor in Carl Corks Biochemistry Department at Washington University in St. Louis. Udenfriend could not imagine, as he and his wife Shirley left for St. Louis, that several of the scientists with whom he had interacted at Goldwater Memorial and New York University, including Shannon, Brodie, Julius AxeIrod, John Burns, and Ochoa, would cross his path again in the years to come. The Department of Biochemistry at Washington Univer- sity, headed by Nobel laureate Carl Cori, was one of the most prestigious biochemistry departments in the country.
SIDNEY UDENFRIEND 273 Udenfriend applied his isotope-derivative methodology to enzymes that were uncler investigation in the Cori labora- tory. One of his close colleagues cluring that period was Sill Velick, en c! their studies resultec! in several papers on amino acid analysis en cl protein N-terminal analysis ~ ~ 95 I, I-2) . On April 7, 1949, the New York Times informal the public on the appointment of James Augustin Shannon as associate director in charge of research at the National Heart Institute created in June 1948 by an Act of Congress signecl by President Truman. This was the beginning of the mete- oric rise of the National Institutes of Health (NIH), of which Shannon became director in 1955, from a routine govern- ment laboratory to the woricl's center of biomeclical science Uclenfriencl received the letter of invitation to join Shannon's expanding research team in 1950 while in St. Louis. His answer when he asker! Cori for acivice was: "If you join a little known government laboratory, this will be the end of your scientific career!" At that time Uclenfriencl also hac! an application pencling for an assistant professor- ship at Columbia University, with little chance of success. So he clicl not hesitate to ignore Cori's acivice en cl accepted the position of biochemist (at the GS-13 level) in the Labo- ratory of Chemical Pharmacology uncler his oIcl boss Broclie in the National Heart Institute, which started in Builcling 3 on the NIH campus in Bethesda, Maryland. By the early 1950s NIH hacl attracted a large group of scientists from GoIc~water Memorial Hospital, in aciclition to Shannon en cl Broclie. At that time NIH was still a flecigling research cen- ter, but the scientific talent present in Builcling 3 in the early 1950s was extraordinary.) In AxeIrod's words: "Never hac! such a small group of promising scientists reacher! such Olympic heights." In a letter ciatecl June 5, 1950, Shannon informal one of us (B.W.) of the complementarily of current projects at
274 BIOGRAPHICAL MEMOIRS Harvarc! University with those of UclenfriencI, en c! so it hap- penecl that Uclenfriencl became a colleague en cl friend un- til his passing. In 1953 one of us (H.W.) was recruited by Uclenfrienc! from his alma mater, CCNY, en c! became his first Ph.D. student, thanks to a graduate program that Uclenfriencl helpecl establish between the Broclie laboratory at NIH en c! the Departments of Biochemistry en c! Pharma- cology at George Washington University. To a young graclu- ate student the quality of science en cl the excitement en cl talent that surrounclec! him in Uclenfriencl's laboratory en c! all of Building 3 left a lasting impression never to be equaled. In this convivial atmosphere at NIH Shannon initiated a weekly interclisciplinary seminar supplemented! by more re- laxecl gatherings of the "Appliecl Statistics Club," a euphe- mism for the poker games with high stakes, where Irish Mist was server! uncler the motto "The Irish never missecI!" lust as Shannon never forgot his famous mentor Homer Smith, so Uclenfriencl acknowlecigecl throughout his scien- tific lifetime that he stool! on the broac! shouiclers of Shan- non. On the occasion of a festive banquet of the Commit- tee for the Weizmann Institute in New York, "godfathers" UclenfriencI, AxeirocI, en c! Witkop cleciclec! to move the au- thorities to name the pilIarecl central administrative builcI- ing, referred to as Building I, the James Augustine Shan- non burbling. After high-level en c! congressional cleliberative clelays a solemn celebration in the presence of a smiling Shannon prececlecl the official christening on January IS, 1983. This was the first en c! unfortunately the last time that an NIH builcling was namecl after a scientist en cl not a mem- ber of Congress. Of the more whimsical talks on this occasion Hans Stetten compared the Shannon building at NIH to the CNS with numerous afferent en cl efferent channels, which Shannon successfully controlled in spotting action potentials amidst
SIDNEY UDENFRIEND 275 much background! noise. Like Ben Franklin he Tookoc! for helpful temperature-clepenclent currents to move the large NIH vessel through stormy seas en cl to avoid unfavorable counter currents the same way as Franklin hac! acivisec! trans- atIantic shipping in 1786 (Transactions of the American Philosophical Society 2 ~ ~ 786) :294-329) . Measuring the "tem- perature" on the "climate of expectancy" in institutes en c! laboratories en cl at the same time respecting their integrity en cl inclepenclence was Shannon's style, en cl therefore, "The style is the man." Hans Stetten later became the first chair of the Scientific Advisory Board to Uclenfriencl at the Roche Institute of Molecular Biology. The transition of pharmacology, based on physiological evaluation, to a science basecl on quantitative analysis using exact colorimetric, fluorescence, or raclioactive-isotope meth- ocis gave Broclie's laboratory the title " chemical pharma- cology" en cl goes back in part to investigations by Uclenfriencl with Keston and Velick. Udenfriend always believed, regardless of the project, that the time best spent was working out a rapid en cl sensitive assay. Here we also have the beginning of research that usecl isotopically labelecl substrates to quan- titatively determine enzyme activity, which lee! to the clis- covery of the famous "NIH shift," as cliscussecl below. During the 1950s hyciroxylation was a common theme in Uclenfriencl's research, en c! it was cluring that perioc! that he became especially interested in aromatic hydroxyla- tion. His first studies on the enzymatic conversion of phe- nylalanine to tyrosine were done with Jack Cooper (1952), and this research soon broadened to include studies on tryptophan hyciroxylation en cl the biosynthesis of both nore- pinephrine en c! serotonin, en c! later proline hyciroxylation and collagen synthesis. He was intrigued by the discovery of serotonin, which was isolated, identified, and crystallized in 1948 by Maurice Rapport in the laboratory of Irvine Page,
276 BIOGRAPHICAL MEMOIRS who was then the director of the Research Division of the Clevelancl Clinic. These collaborative studies clevelopecl into both personal en cl productive relations between the two groups. The first step in the serotonin biosynthetic pathway stucliecl in cletail was the conversion of 5-hyciroxytryptophan (5HTP) to serotonin ~ ~ 953, I, ~ 954, ~ ~ At first it was thought that this enzyme was distinct from the clecarboxylase that usecl clihyciroxy-phenylalanine (DOPA) as substrate, but upon purification the enzyme, callecl aromatic amino acid clecar- boxylase, was shown to be able to clecarboxylate not only 5HTP and DOPA but also tryptophan, tyrosine, and pheny- lalanine, although to a lesser extent. By ~ 953 it became clear that serotonin biosynthesis in- volvecl two steps, hyciroxylation to 5HTP en cl clecarboxyla- tion to serotonin. By then it was also apparent that seroto- nin was not only a neurotransmitter but hac! a role as a vasoconstrictor en cl potentially other roles because of its high concentration in both platelets en cl intestinal mucosa. This surge in the central en c! peripheral importance of se- rotonin lecl to extensive basic en cl clinical investigations in which Uclenfriencl en cl his colleagues or clisciples, such as Herb Weissbach, Walter Lovenberg, ETwood Titus, and the clinical group heaclecl by Albert Sjoercisma, were involved. Carcinoicl syndrome is just one example of the productive collaboration between the Uclenfrienc! en c! Sjoercisma group. These tumors produce large amounts of serotonin that cause the gastrointestinal symptoms en cl blushing seen in these patients. Weissbach had already developed an assay for 5- hydroxyindole acetic acid (5HIAA), the primary urinary metabolite of serotonin. Thus a simple diagnostic test for the malignant carcinoic! syndrome was clevelopec! baser! on the determination of 5HIAA in urine (1955,1~. An interest- ing siclelight to these studies was the observation that Weissbach was routinely running high levels of 5HIAA in
SIDNEY UDENFRIEND 277 his urine while others in the lab hac! normal levels. There, of course, was concern that he might have a carcinoicl tu- mor until the high 5HIAA levels were tracecl to his ciaily ingestion of bananas that contain high levels of serotonin en cl other amines. This work was extenclecl to other fruits en cl vegetables, which brought Uclenfriencl into contact with nonscientists like the president of the Uniter! Fruit Com- pany, which lecl to an award to Uclenfriencl en cl Sjoercisma sponsored by Unitecl Fruit. The neec! to localize en c! assay serotonin was one of the reasons that Robert Bowman, the chief of the Laboratory of Technical Development, helpecl Sill to design a spectro- fluorometer (SPF) with quartz optics that not only extenclec! fluorescence assay into the ultraviolet region but also per- mittecl one to change both the activation en cl fluorescent wavelengths to achieve increaser! sensitivity en c! much higher specificity (1955,2~. The initial instrument, put together by Bowman using some parts from an Army en cl Navy store in Bethesda, took up half a laboratory en c! because there was no shielcl to prevent room light from activating the SPF photomultiplier, the room hacl to be kept dark cluring the measurements. Using this instrument the sensitivity of the serotonin assay increased by orders of magnitude and it was now possible to assay enclogenous serotonin in virtually any tissue (1955,3~. This ciramatically changer! the research ef- forts en cl opened up a new dimension in biogenic amine research. The clevelopment by the Aminco Company of a small well-clesignec! SPF (caller! the Amino-Bowman SPF) also macle it possible for the scientific community to have access to this new instrument. Numerous assays were clevel- oped for all sorts of compounds using the SPF as described in the book Uclenfriencl first publishecl on fluorescence as- say in biology en cl medicine in 1962, with a second eclition in 1969. How this story evolves! in 1955 is also clescribec! by
278 BIOGRAPHICAL MEMOIRS Uclenfrienc! in a nostalgic retrospection 40 years later pub- lishecl in Protein Science (4~19951:542-51~. In a surprising about-face the mentor-clisciple role with Siciney Velick was reverser! when the two Sicis collaborates! on the use of the SPF on novel en cl previously inaccessible problems such as enzyme-co enzyme complexes or antigen-antibocly in terac- hons. Several Nobel Prize winners have reliecl on the SPF as an inclispensable tool. In collaborative studies AxeIrocl iclen- tifiec! labile metabolites of lysergic acic! cliethylamicle (LSD), mescaline, en cl norepinephrine. In AxeIrocl's worcis, The SPF made it possible to measure noradrenaline and serotonin . . . practically. This changed the direction of the whole field of neurobiology. Quantitive studies established the relationship of the level of these trans- mitters to certain mental illnesses and aided in the development of mental tranquilizer and energizer drugs. Continued studies in this area will yield additional information on the basis for mental illness. The aciage "Transmission is as important as discovery" couIcl be appliecl to the time that Uclenfriencl spent as a graduate student with Ochoa in the Department of Bio- chemistry at NYU Meclical School in 1946. Uclenfrienc! be- came aware that hydroxyproline was uniquely present in collagen from his earlier clays at NYU, since Joseph Bunim, a professor at the NYU Meclical School, hac! impresser! on him how collagen was intimately involvecl in the health en cl disease of connective tissue, in arthritis en cl other clisor- clers. Bunim en c! Stetten soon joiner! NIH at the Institute of Experimental Biology, which was not accepted by Con- gress as a serious "disease" en cl so became the National Institute of Arthritis en c! Metabolic Diseases (NIAMD). That the hyciroxylation of proline floes not occur in the free form, but at some step in the formation of collagen was the discovery of Marjorie ("Marnie") Stetten en c! in-
SIDNEY UDENFRIEND 279 spires! UclenfriencI, aireacly involves! in hyciroxylation reac- tions, to pinpoint the exact step at which proline was hy- ciroxylatecI. Uclenfriencl hacl the goocl fortune of having Beverly Peterkofsky join the laboratory at that time. Peterkofsky, a graduate student with Ochoa at NYU, movecl to NIH when her husband Alan Peterkofsky accepted a po- sition in the NIAMD. As Uclenfrienc! sail! of Beverly Peterkofsky, "It was one of the best things that ever hap- penecl to me." She finished her graduate studies in Uclenfriencl's lab, where she obtainer! a cell-free system from chick embryos ~ ~ 96 ~ ~ that incorporated cis-and trans-4-H3- L-proline into pepticle-bouncl hyciroxyproline in a front-sicle clisplacement with complete retention of configuration at C-4 (1964~. This reaction was comparable to other enzymes, which clirectly use molecular oxygen in the formation of hydroxylated products. Years later, in 1975, at a Collagen Symposium at the Roche Institute of Molecular Biology, Uclenfriencl fancily remembered these early events in the collagen saga that was completer! by Darwin Prockop, an- other of Uclenfriencl's students. A major discovery was the fincling that alpha-ketoglutarate was the cofactor of proline hyciroxylase en c! of a totally new class of enzymes. This en- ablecl Prockop to study in cletail the nature of the hyciroxy- lase en cl of the transformation of "protocolIagen" into col- lagen. Car! Piez at NIH then carrier! out a kinetic stucly of collagen biosynthesis, en cl Prockop showocl a role for hy- ciroxyproline in stabilizing the triple helix of collagen that Lubert Stryer in his famous textbook Bio-Chemistry likenec! to a Bach fugue. Prockop continual his studies on collagen after leaving the Uclenfriencl lab en cl moving to Philaclel- phia, where he shower! that mutations in the genes for col- lagen, caused osteogenesis imperfects, or brittle bone dis- ease in chilciren, or dwarfism (chonciroclisplasias), not to mention the role of collagen in more common syndromes,
280 BIOGRAPHICAL MEMOIRS such as osteoporosis en c! osteoarthritis. These insights rounc! out the clinical observation by Uclenfriencl en cl Sjoercisma of the increased excretion of hyciroxyproline in Marfan's syndrome, which goes back to 1958 en c! even further when we consider Egypt's eighteenth dynasty with Amenopsis en cl Tutankhamen being possible victims of this clisorcler. As early as 1953 Uclenfrienc! en c! Samuel Bessman pub- lishecl on the hyciroxylation of phenylalanine in patients with the genetic disease pheny~pyruvic oligophrenia, called phenylketonuria or PKU (1953,2~. This work prececlec! the exhaustive research on phenylalanine hyciroxylase by NIH colleague en cl friend Seymour Kaufman that extenclecl over 20 years. A mocle! system for aromatic hyciroxylation pub- lishecl in 1954 with Broclie en cl AxeIrocl was intenclecl to throw some light on the mechanism of this oxidation (1954,2~. Witkop informer! Uclenfrienc! that his system con- sisting of oxygen, ferrous ion, en cl ascorbic acicl in the pres- ence of ethylenecliaminetetracetic acicl (EDTA) is a moclifi- cation of a system that Heinrich Wielanc! clescribec! for the oxidation of formic acicl by ferrous ion, clihyciroxymaleic acicI, an agent forming metal complexes en cl oxygen, as mentioner! in his Silliman memorial lectures (Yale Univer- sity Press, 1932, p. 86~. "Progress is tradition preserved." Curiously enough this rediscovery went into the literature as "Uclenfriencl's reagent" (Michael B. Smith, Organic Syn- thesis, McGraw-Hill, 2002, p. 296~. Here we deal with the name game (Alex Nickon, Modern Coinec! Terms en c! Their Origins, Pergamon Press, ~ 987) to which we will return subsequently. There is a similarity of this Wieland-Udenfriend system with the requirements of proline hyciroxylase for alpha-ketoglutarate, ferrous ion, ascorbate en c! oxygen as founcl in ~ 966 ~ ~ 966, ~ ~ . The years ~ 966 en cl ~ 967 were the time when insight into the mechanism of hyciroxylation was obtained because of the availability of a tritiated substrate,
SIDNEY UDENFRIEND 28 p-H3-phenylalanine, for phenylalanine hydroxylase, the im- portant enzyme missing in PKU. When the results of the experiment on the fate of the tritium came in, Uclenfriencl was perplexed, for there was much tyrosine former! but no loss of tritium. Questioning the location of the tritium in the commercial sample, Sicl angrily scolclecI, "Can't chem- ists put the label in the right place?" In discussing this cli- lemma with Gordon Guroff of Uclenfriencl's laboratory, John Daly, a chemist in Witkop's laboratory, macle the sugges- hon of extending the stucly to cleuterophenylalanine, where NMR could rigorously establish the position of the cleute- rium. Using this as substrate for the enzymatic hyciroxyla- tion some cleuterium was lost but most was fount! in the meta-position, as was then shown for tritium (1967,1~. This was the birth of the happy child christened the "NIH shift," with several prouc! parents involves! in paternity (1967,1~. There even was a twin: Not only clicl tritium en cl cleuterium slicle over to the neighboring meta-position but so clicl halo- gen (1966,2~. Of course, tryptophan-5-hydroxylase, the first step in serotonin synthesis, was the next enzyme to be testecI.2 The reaction hacl only been clemonstratecl in whole cells of Chromobacterium v~o~accum. Using as substrate 5-tritio-tryp- tophan, 4-tritio-5-hydroxytryptophan was formed with little release of tritium into the meclium (1966,3~. To strengthen the case for a postulated arena-oxide in- termecliate3 Donald farina, a chemist in Witkop's labora- tory, synthesized benzene oxide, and naphthalene I,2-ox- icle, using the synthetic methods of Emanuel Vogel, the pioneer in the arene oxide field. The latter oxide was iclen- tifiecl as an intermediate cluring conversion of naphthalene to 2-naphthol en c! transnaphthalene-1,2-clihycirocliol by the action of liver enzymes ~ ~ 970) . The hydroxylation of the antipyretic and anti-rheumatic acetanilicle to the more active p-hyciroxy-metabolite acetami-
282 BIOGRAPHICAL MEMOIRS nophen, later soil! as Tylenol, was a discovery of Axeiroc! in Broclie's laboratory. AxeIrocl often mentioned that he missecl becoming a millionaire many times over by not patenting this process. When this same process was reinvestigates! with p-tritioacetanilicle, radioactive "Tylenol" was formecl both in vitro with microsomes en cl in viva (1972~. The migration en c! retention of tritium ranger! between 38 percent en c! 56 percent. Nonenzymatic hyciroxylations of aromatic substrates leacl to the NIH shifts only with peroxytrifluoroacetic acicI, a much stronger oxidant than the Uclenfrienc! reagent. Uclenfriencl always hacl close contact with Seymour Kety, because they both triecl to final the biochemical basis for mental clisorclers, a fact that lee! to the first International Symposium on Catecholamines at NIH in October 1958. Uclenfriencl en cl Witkop presented there the ooservanon on the conversion of clopamine to 6-hyciroxyclopamine, which selectively destroys catecholamine-containing nerve termi- nals en cl was at one time thought to be a possible enclog- enous agent involves! in mental diseases. Sill hacl an unfailing eye for bucicling talent, en cl it is not possible to document the large number of successful scientists who passer! through his lab or the impact he hac! on so many others. One such example is the case of Paul ~ '.r., ~ , ~ , ~ GreengarcI, who came to Uclenfriencl's laboratory in the mid-1950s to learn assays and some of the procedures be- ing routinely clone in the amine fielcl before beginning a position at Ciba-Geigy. Greengard studied the uptake of tyrosine in the rat brain, a beginning that he gratefully remembered when he received the Nobel Prize in 2000 for extending this initial interest in the brain to highly refined receptor studies. The fact that Marshall Nirenberg, who received the Nobel Prize in 196S, remained at NIH after his initial experiments in the early ~ 960s that cracker! the genetic cocle was in
SIDNEY UDENFRIEND 283 large part clue to Uclenfriencl's efforts, en c! the coincidence that Nirenberg's wife, Perola, was Uclenfriencl's assistant. Weissbach remembers clearly when Uclenfriencl callecl a lab meeting to tell us that there was a chance that Nirenberg wouIcl leave NIH unless he hacl more space to continue his experiments: "We all agreed to cooperate, en cl soon there- after Nirenberg's group mover! into the space we macle avail- able. I benefited greatly from the proximity of the Nirenberg group en cl within a short period of time was actively en- gagec! in experiments to eTuciciate how the genetic infor- mation was usecl in the translation process." Marshall Nirenberg sums up his memories as follows. My wife, Perola, worked as a technician for Sid Udenfriend for about ten years, from 1958 to 1968. Perola had enormous admiration for Sid and he valued her work greatly. Theirs was probably the best working relationship I have ever seen. One of the reasons why it was so successful is that Sid would outline a problem to Perola and suggest a possible mode of attack and Perola then would set up the assays and see if it would work. After a month or so when she had some data she would go back to Sid and show him the data or discuss problems she had encountered. So Perola had all of the fun of solving most of the problems she encountered on her own, and Sid could do exploratory research while investing very little of his own time. It was an ideal arrangement for both of them. One day Perola asked Sid for a few days leave so that she could go with me to visit an academic institution that had offered me a position, and she told him that I probably would accept the position. By the time we returned to Bethesda Sid had worked out a plan to keep me at NIH by offering me some space and support within his laboratory, which would enable me and my colleagues to continue our work. And so I moved to Sid's lab. Years later Sid often enjoyed telling me that the reason that he had offered me the position in his lab was to keep Perola from leaving NIH, and I would counter by saying that he was just plain lucky to have gotten me to go to his lab. In fact, this arrangement proved mutually beneficial because Sid and
284 BIOGRAPHICAL MEMOIRS Perola continued to work with one another, and my colleagues and I were able to finish deciphering the genetic code. Our presence in Sid's lab made it easy for Herb Weissbach to begin working on protein synthesis, since we were experts in the field. Sid Udenfriend always was bubbling over with enthusiasm and ideas for the projects that he was involved in. He had a superb mind and would have been successful in almost any field of endeavor. He always tried to help me in a fatherly way by giving me the benefit of his own experience. Sid called me a few weeks before his death to find out how Perola and I were, and tell me about his plans for the future. He especially wanted me to convey his best wishes to Perola. I think that Sid was an outstanding human being as well as an outstanding scientist. Uclenfriencl's career was flourishing at NIH en cl by the early 1960s he was continually being approached about po- sitions in both academia en cl industry. With only a few ex- ceptions he expressed little interest in leaving the woncler- fuT, stimulating environment in Bethesda. It wouic! take a unique challenge to pull him away from this research Mecca, en cl in 1967 such an opportunity appeared, clue in large part to oic! friendships. John Burns, a former colleague of Uclenfriencl's from the GoIc~water Memorial period, hacl movecl his laboratory to NIH in 1957. Burns remained there for only a short perioc! en c! then became vice-presiclent o research at Burroughs WelIcome in 1960. In January 1967 he movecl to Hoffmann La Roche as vice-presiclent of re- search and met Udenfriend at a cocktail party in Bethesda shortly after assuming his post at Roche. Burns was anxious to make innovative changes in Roche research, and Uclenfrienc! suggester! that Roche establish a basic science institute as part of the company's research effort. Unlike existing programs at most pharmaceutical companies this institute wouic! not be product driven but function much like the intramural NIH, with the scientists having direct If
SIDNEY UDENFRIEND 285 funcling, a reasonable time commitment, en c! freedom to pursue a research project of their own choosing. The ben- efits to the company wouIcl come from the cutting-ecige research that wouic! place the company in a unique posi- tion to move rapicIly into new areas of biology en cl clevelop novel therapeutics. It was becoming clear even by 1967 that the discoveries in molecular biology en c! molecular genet- ics that Uclenfriencl was so aware of because of his associa- tion with the Nirenberg laboratory, wouIcl be the ciriving force for the clevelopment of new cirugs in the clecacles to come. From this brief casual discussion at a social gathering arose the concept of the Roche Institute of Molecular Biol- ogy (RIMB). Within the short span of four months, thanks to the efforts of UclenfriencI, Herb Weissbach (whom Uclenfrienc! hac! asker! to join him), en c! Burns, the RIME came into being. By April Burns presented a summary pro- posal en cl cletailecl buciget to the Roche Executive Commit- tee. Approval from Nutley en c! Base! came quickly, thanks in part to Alfrecl Pletscher, who was heacl of research in Roche Basel. Pletscher hacl spent time in the micI-1950s in Udenfriend's laboratory working directly with Weissbach and was supportive of the concept. IncleecI, clue to the efforts of Pletscher, within two years the Basel Institute of Immunol- ogy, the sister institute to the RIME, was establisher! in Base! near the Roche facilities. The period between May en cl July 1967 was a critical time in the history of the Roche Institute. UclenfriencI, Weissbach, en cl others were unsure whether a move to in- clustry, despite the attractiveness of what was being plannecI, was too big a career risk to take. At that time basic scientists were extremely wary of industry. It was clear that Udenfriend wouIcl not make the move without a solicl contingent of committed scientists. With Shannon the man and his talent
286 BIOGRAPHICAL MEMOIRS came first en c! then the mission. In this way he assembler! the stellar cast that lecl NIH to such scientific success in the same way as UclenfriencI, after he movecl to Roche, hacl the satisfaction of assembling a similar group. Whether Roche wouIcl keep its promise to establish en cl maintain a basic research institute for a reasonable period (e.g., a 10-year commitment) was the major question. Finally a meeting was scheclulecl in June of 1967 with V. D. Mattia, then presi- clent of Roche in Nutley. A group of scientists from NIH that Uclenfrienc! wan tee! to recruit, all with great concern, met with Mattia. By the end of the meeting it was clear that the title hacl turned. Although a time period was never put in writing, the scientists came away convincer! that the Roche commitment was long-term en cl most of the scientists at that meeting eventually joined the RIMB. Shortly thereaf- ter the freedom the scientists clesirec! wouic! be clearly states! in a charter signecl by Mattia on July 14, 1967. That was the clay the Roche Institute of Molecular Biology came into being. The RIMB laster! 28 years en c! cluring its existence the commitment that Mattia macle to the NIH scientists in 1967 was never broken. Although Mattia passed away be- fore construction of the institute was finisher! in 1971, suc- ceecling presidents, such as Robert Clark en cl Irwin Lerner, respected the provisions in the RIMB charter. Once the charter was in place events mover! quickly. Within months Udenfriend obtained commitments from a number of young NIH scientists, including Herb and Arthur Weissbach, Nathan Brot, Sydney Spector, Sidney Pestka, Ronalcl Kaback, en cl Aaron Shatkin. Richard Snycler was hired to hancIle the administrative affairs, en cl temporary office space was renter! in Be thescia. A clistinguishec! Bo arc! of Scientific Advisors was established and by the summer of 1968 temporary space was available in Nutley and Udenfriend en c! scientists in his department set up the first laborato-
SIDNEY UDENFRIEND 287 ries. In 1971 Uclenfrienc! was electec! to the National AcacI- emy of Sciences, which gave prestige to both the RIMB and the company. By 1971 construction of the Roche Institute was completecI, ant! the RIMB scientists who hac! been houses! in temporary laboratories throughout Roche en cl in labora- tories across the nation en cl abroacl were able to move into the new buckling. For the first time the institute staff was together uncler one roof. Uclenfriencl was icleally suited to be director of a basic research center serving the pharmaceutical industry. Although the scientists in the institute hacl free rein, Uclenfriencl hacl the unique ability of seeing a practical application to many of the programs. In this way the company always had a direct line to what was happening in the institute en cl the opportunity to have the technology transferred without in- terfering with the research philosophy the institute was built on. Two of the initial members of the RIMB who Uclenfriencl had brought from NIH made important discoveries early on that were of interest to the parent company. Syciney Spector developed an assay for drugs of abuse, which be- came a major product of Roche Diagnostics, en c! Sidney Pestka, whose work on interferon brought Roche into the fielcl of biotechnology, were clear examples of how the con- cept of a basic research institute within a pharmaceutical company could be successful. Under Udenfriend's leader- ship the environment at the Roche Institute was conducive to cloing goof! science en c! the careers of many of the scien- tists flourished there. Basecl on the work clone at the RIMB, three of the members, Aaron Shatkin, Herb Weissbach, en cl RonaTc! Kaback, were elected! to the National Academy of Sciences en cl at one point the RIMB hacl seven members of the Academy, inclucling Severo Ochoa, Bernard Horecker, en c! Allan Conney, among a staff of less than 30 scientists.
288 . BIOGRAPHICAL MEMOIRS In aciclition, the RIMB hac! a training mission. From the initial discussions in 1967 it was clear that the long-term success of the RIMB as a basic research center wouIcl cle- penc! on being able to attract postcloctoral fellows en c! graclu- ate students. Uclenfriencl was cleterminecl that this wouIcl be the case. The charter clearly statecl training as a mission of the RIMB. At that time, in the late 1960s, universities were reluctant to accept industry scientists (as the institute scientists were viewocI) as adjunct faculty en cl there was a perioc! of great concern for Uclenfrienc! that institute scien- tists wouIcl not have university affiliations en cl thus not be able to have graduate students. UclenfriencI, thanks to oIcl friendships with faculty members at City College, such as Abe Mazur en cl Mike Fishman, obtained the first appoint- ment from his alma mater, City College. Within a short perioc! a strong relationship was built between the RIMB en cl Columbia University, thanks to the efforts of Uclenfriencl en cl Sol Spiegelman who was the new chair of the Depart- ment of Human Genetics at Columbia. Eventually RIMB scientists hacl appointments at most of the large universities in the New York-New Jersey region. Postdoctoral fellows were anxious to come, en c! there was no aspect of the RIMB that Uclenfriencl was more prowl of than the fact that through the 28 years the RIMB was in existence more than 1,000 postcloctoral fellows en c! close to 50 graduate students re- ceivecl their training at the RIMB. Uclenfriencl's own research never falterecl cluring the perioc! he was director from 1968 to 1983. He continues! his studies on the hydroxylation of proline, tyrosine, and clopamine (1971,1-2, 1972~. In aciclition to a basic interest in the mechanism of these reactions, Uclenfrienc! always consiclerecl the in viva ramifications en cl attempted to un- clerstancl how proline hyciroxylase was involvecl in collagen synthesis and how tyrosine hydroxylase and dopamine beta-
SIDNEY UDENFRIEND 289 hyciroxylase were involves! in the regulation of norepineph- rine synthesis. During this period his love en cl knack of cleveloping assays lecl to the use of fluorescamine as a sensi- tive reagent for the assay of amino acids, peptizes, en c! proteins (1973~. The clevelopment of the fluorescamine as- say macle it possible to detect small amounts of peptizes en c! proteins cluring purification en c! was especially valu- able in Uclenfriencl's studies on the enkephalins, opioicl peptizes, as well as the separation en cl isolation of various species of natural oc-interferon by HPLC. The studies on interferon deserve special attention, for this was the first example of how research in molecular biology prover! valuable to the company. The interferon project was initiated at RIMB by Pestka, who felt that this naturally occurring protein might have both antiviral en cl antitumor activity. In order to clone the gene for this chemokine it was necessary to purify it first from white bloocl cells en cl obtain a partial amino acid sequence. The lack of large amounts of cells en c! the realization that there may be a family of interferons macle the task much more clifficult. Without the analytical procedures that were available in Uclenfriencl's laboratory it is cloubtfuT that the isolation of the first natural interferon species wouIcl have been achieved so quickly. Once the purification was achieved the sequenc- ing en c! cloning of an interferon gene was accomplishes! in Pestka's laboratory. By the early 1980s, in collaboration with scientists at Genentech, oc-interferon became the first Roche cirug proclucec! by recombinant DNA technology. It server! as a prototype for other biotechnology products (e.g., interIeukin-2), and it is well accepted that the RIMB was the prime factor in making Roche one of the first, if not the first, large pharmaceutical companies to move into bio- technology. The influence that the RIMB hacl on the course of Roche research was living proof of Uclenfriencl's vision
290 BIOGRAPHICAL MEMOIRS of the role of the institute when it was first conceiver! in 1967. In 1983 at the age of 65, Uclenfriencl stepped clown as director of the RIMB, en c! the reins were passer! to Herb Weissbach. UclenfriencI, of course, was not really to retire en cl continual to direct a productive laboratory. His pri- mary research cluring the late 19SOs en c! early 199Os cen- terecl on alkaline phosphatase en cl its attachment to the cell membrane by a phosphaticlylinositol containing a gly- colipic! anchor. Uclenfriencl's studies helpec! to eTuciciate the biogenesis of this unioue linkage. cleavage. and oro- cessin~ of the anchored proteins. - -a ~ - -a - Like the other scientists at the RIMB, many of them younger aciclitions to the staff, he was looking forward to productive years at the Roche Institute. However, Hoffmann- La Roche, although one of the major large pharmaceutical companies in the world, was facing financial constraints that were initially apparent after the expiration of the Valium patent in the early 19SOs. By 1994 major Tong-term cleci- sions were being macle about the future direction of the company research, en cl to the surprise of the RIMB staff Weissbach, who was director at the time, was informer! that the RIMB wouIcl be phasecl out. Weissbach hacl the un- pleasant task of terminating the institute in a manner that was least destructive to the institute staff. For both Weissbach en cl Uclenfriencl this was the most clifficult period in their long careers. What they hacl started together almost 30 years ago was coming to an end. Weissbach worked with Roche management to insure that all of the members of the insti- tute would leave with their equipment, as well as some sup- port if they were moving to a university position. It took almost two years for everyone in the institute to be placecI. At times Udenfriend found it difficult to deal with the clismantTing of the institute, which hac! meant so much to
SIDNEY UDENFRIEND 29 him, although he en c! Weissbach kept in touch cluring the long negotiations. By December 1995, about a year after the initial announcement of the closing of the RIME, most of the institute staff hac! left. Weissbach hac! cleciclec! he wouIcl not leave until everyone was placecI, en cl still main- tainecl a functioning laboratory. He wouIcl soon leave the institute buckling, which was being closecI, en c! move to another location within Roche. Dreaciful as the closing of RIME was for UclenfriencI, in December of 1995 he wouIcl face a major unexpected! challenge that wouic! obscure all other concerns. Early in that month Uclenfriencl en cl his wife, Shirley, hac! stopper! at a pharmacy in Ceciar Grove, New Jersey, to pick up a prescription. He had parked facing a brick wall en cl put the car in reverse as he was preparing to return home. What happened! next is still not clear. It appears that when the car was put into drive, it acceleratecl rapicIly en cl crashed into the brick wall some 30 feet in front of the car. Both Uclenfrienc! en c! Shirley sufferer! multiple fractures, en cl Uclenfriencl was in a coma for several clays after the accident. Although both wouIcl survive the accident, in that one split seconc! their lives were irreversibly changed. After months of rehabilitation they both were finally able to re- turn to their home. Weissbach hacl set aside an office for Uclenfrienc! in his new space at Roche, en c! Uclenfrienc! wouIcl come in about once a week, more to chat with Weissbach than to clo science. By the fall of 1996 everyone in the institute hac! been placecI, en c! Weissbach was plan- ning on closing clown his laboratory in December en cl relo- cate to a position at Floricia AtIan tic University. The equip- ment was being moved on a dreary, damp Saturday in December of 1996, and Weissbach, there alone, was un- aware that Udenfriend had made it a point to come in that day, since this was the last day of the RIMB. Weissbach did
292 BIOGRAPHICAL MEMOIRS not have to ask Uclenfrienc! why he hac! botherec! to come. Uclenfriencl's first worcis were, " We started this institute together en cl I wanted to be here when it enclecI." By noon the two left the buckling in the freight elevator, through the loacling clock. They realizecl that for the first time in more than 40 years their career paths wouIcl diverge. The reasons for the clemise of the Roche Institute of Molecular Biology are still not entirely clear. The end of this worIcI-renownecl research center that housecl so many outstanding investigators touches at the root of the reasons for research support that perhaps was expressed nowhere better than by Arthur Kornberg (Nobel Prize, 1959~. The difficulty with research support in our society, I have come to realize, is the failure to understand the nature and importance of basic research. This failure can be seen among members of the lay public, political lead- ers, physicians, and even scientists themselves. Most people are not pre- pared for the time-scale of basic research and the need for a critical mass of collective effort. Fragments of knowledge [unwelcome] and unexploited are lost, as were Gregor Mendel's basic genetic discoveries. The vast major- ity of legislators and some scientific directors cannot accept the seeming irrelevance of basic research. Were there a record of research grants in the Stone Age, it would likely show that major grants were awarded for propos- als to build better stone axes and that critics of the time ridiculed a tiny grant to someone fooling around with bronze and iron. People do not realize that when it comes to arguing their case for more funding, scien- tists who do the basic research are the least articulate, least organized, and least temperamentally equipped to justify what they are doing. In society where selling is so important, where the medium is the message, these handicaps can spell extinction. Uclenfriencl was an outstanding researcher en cl teacher but perhaps his greatest contribution to science was in es- tablishing the Roche Institute of Molecular Biology, en c! cluring his tenure as director, in creating one of the out- stancling inclustry-supportecl biological research institutes in the woricI. The success of the Roche Institute is not mea-
SIDNEY UDENFRIEND 293 surer! only by the papers publisher! or the accomplishments of the incliviclual scientists or the impact on the company. What will be its greatest legacy is the large number of indi- vicluals trainee! at the institute, scatterer! throughout the worIcI, who remain to this clay a living reminder of the Roche Institute. Uclenfriencl's ciream hacl come true. When Uclenfrienc! left Roche in December 1996, he al- reacly hacl accepted a position at Drew University as clirec- tor of the Charles A. Dana Research Institute for Scientists Emeriti (RISE). This institute was specifically establisher! to encourage interaction between some of the top retired sci- entists from inclustrv in New Tersev en cl unclergracluate stu- , ~ , clents at the university. Uclenfrienc! remainec! in that post through 1999, en cl uncler his leaclership the institute ex- panclecl its membership en cl broaclenecl its sphere of activi- ties. Uclenfrienc! obtainer! great satisfaction from working closely with unclergracluate students en cl his caring for both science en cl people were apparent to all who knew him at Drew University. In 1999 Uclenfriencl macle the clifficult decision to step clown as director of the RISE. He en cl Shirley hacl cleciclecl to move to Atlanta, where their daughter livecI, since it was becoming clear that because of age en cl the aftereffects of the accident, they both neeclecl help to carry on many of their tinily activities. The move south was macle in 1999, but soon after they were settlecl Uclenfriencl was showing symp- toms of coronary artery blockage. In the early winter of 1999 he entered the hospital for a bypass operation, which appeared to be successful, however, cluring recovery he ap- parently suffered a massive stroke en cl remained in a coma for several days until his death on December 29, 1999. The funeral was helcl on December 3l, en cl because of the time factor en cl location, only about 20 people, mostly his close relatives, attenclec! the gravesicle service. Weissbach was able
294 BIOGRAPHICAL MEMOIRS to fly up from Floricia en c! was the only scientific colleague from the past to be present. Weissbach plannecl on having a memorial event in Uclenfriencl's honor for the many scientists whose lives Uclenfriencl touched. Working with Ashley Carter, the new director of the RISE en cl Barbara Petrack a RISE member, a half-clay symposium was hell! on May 25, 2000, on the Drew University campus. A scientific lecture was presented by GreengarcI, en cl the list of scientific colleagues who macle short remarks, in aciclition to the organizers, incluclec! Witkop, Burns, Nirenberg, AxeIrocI, Spector, Arthur Weissbach, Sjoercisma, Ron Kuntzman, en cl Fishman. Aliza, Uclenfriencl's daughter, was also present. Udenfriend leaves a scientific legacy that includes close to 500 publications en cl major contributions to the fielcis of analytical biochemistry, fluorescence, hyciroxylation reactions, serotonin en cl norepinephrine biosynthesis en cl metabolism, collagen biochemistry, encephalins, amino acid transport, and protein anchoring to membranes. Although research en cl not formal teaching was the focus of his career he trained clozens of postcloctoral fellows, through his univer- sity appointments at George Washington University, City College, en cl Columbia University, among others, he trained a large number of graduate students. His role in establish- ing the Roche Institute was a major accomplishment, but what will be missecl most is the enthusiasm en cl love of science that were an integral part of his being. Sic! Uclenfrienc! is gone but not forgotten. NOTES 1. Included in this list are Nobel laureates Chris Anfinsen, Julius Axelrod, and Arthur Kornberg. Several scientists from that early permanent staff in Building 3 later were members of the National Academy of Sciences: Bruce Ames, Robert Berliner, Donald Fredrickson,
SIDNEY UDENFRIEND 295 Leon Heppel, Bernard Horecker, Earl and Theresa Stadtman, Herbert Weissbach, Bernhard Witkop, and Tames Wyngaarden. Fredrickson and Wyngaarden eventually became directors of NIH. Other out- standing postdoctoral fellows and visiting scientists who worked in Building 3 at that time included Paul Stumpf, Horace Barker, Gerard Hurwitz, Paul Marks, and Arthur Weissbach. The authors realize that this is a partial list and apologize to the many talented scien- tists who worked in Building 3 but have not been mentioned. 2. A tryptophan research meeting on a regular international ba- sis was eventually organized in 1971, mainly as a result in the grow- ing interest in the role of serotonin in depression and moods and the wider consequences for neurochemistry, psychiatry, cardiovas- cular studies, and more recently immunobiology and neuro- immunobiology. The acronym for these biannual symposia is ISTRY, or International Study Group for Tryptophan Research. 3. The First Symposium on Arene Oxides in Biochemistry and Metabolism ~ Science 178~19724:779-81) was held atRochein April 1972 with Udenfriend presiding and pointing out that as early as 1947, E. Boyland, who was present, had postulated arene oxides as reactive intermediates in the metabolism of polycylic aromatic sub- strates, an immense area of research for the carcinogenic effects of tobacco smoke and benzopyrene keeping investigators, such as Harry Gelboin (NIH), Allan Conney, (Roche), Don Jerina, (NIH), Charles Heidelberger (University of Wisconsin), and many others busy for years.
296 BIOGRAPHICAL MEMOIRS SELECTED BIBLIOGRAPHY 1943 With B. B. Brodie. The estimation of quinine in human plasma with a note on the estimation of quinidine. 7. Pharm. Exp. Ther. 78:154- 55. 1949 With A. S. Keston and R. K. Cannan. A method for the determina- tion of organic compounds in the form of isotopic derivatives. I. Estimation of amino acids by the carrier technique. 7. Am. Chem. Soc. 71:249-57. 1951 With S. F. Velick. Isotope derivative analysis for praline, valine, methionine, and phenylalanine. 7. Biol. Chem. 190:721-31. With S. F. Velick. The isotope derivative method of protein amino end-group analysis. 7. Biol. Chem. 190: 733-40. 1952 With J. Cooper. The enzymatic conversion of phenylalanine to ty- rosine. J. Biol. Chem. 194:503-11. 1953 With C. T. Clark and E. O. Titus. A new route of metabolism of tryptophan. 7. Am. Chem. Soc. 75:501. With S. Bessman. The hydroxylation of phenylalanine and antipy- rine in phenylpyruvic oligophrenia. J. Biol. Chem. 203:961-66. 1954 With C. T. Clark and H. Weissbach. 5-Hydroxytryptophan decar- boxylase: Preparation and properties. 7. Biol. Chem. 210:139-48. With B. B. Brodie, J. Axelrod, and P. A. Shore. Ascorbic acid in aromatic hydroxylation. II. Products formed by reaction of sub- strates with ascorbic acid, ferrous ion and oxygen. 7. Biol. Chem. 208:741-50. With K. Kodukula, D. Cines, and R. Amthauer. Biosynthesis of phosphatidylinositol-glycan (PI-G) membrane anchored proteins
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298 BIOGRAPHICAL MEMOIRS ing enzymatic hydroxylation of p-deutero- and p-tritioacetanilide. Arch. Biochem. Biophys. 120:413-19. 1968 With J. W. Daly, G. Guroff, and B. Witkop. Hydroxylation of alkyl and halogen substituted anilines and acetanilides by microsomal hydroxylases. Biochem. Pharmacol. 17:131-36. 1970 With D. M. Terina, T. W. Daly, B. Witkop, and P. Zaltzman-Nirenberg. The role of arene oxide-oxopin system in the metabolism of aro- matic substrates. III. Formation of 1,2-naphthalene oxide from naphthalene by liver microsomes. 7. Am. Chem. 90:6525-27. 1971 With G. T. Cardinale and R. E. Rhoads. Simultaneous incorporation of i8O into succinate and hydroxyproline catalyzed by collagen praline hydroxylase. Biochem. Biophys. Res. Commun. 43:537-43. With W. Dairman. Decrease in adrenal tyrosine hydroxylase and increase in norepinephrine synthesis in rays given L-dopa. Science 171 :1022-24. 1972 With B. K. Hartman and D. Zide. The use of dopamine-a-hydroxy- lase as a marker for the central noradrenergic nervous system in rat brain (immunofluorescence/microcirculation/norepinephrine) . Proc. Natl. Acad. Sci. U. S. A. 69:2722-26. 1973 With S. Stein, P. Bohlen, T. Stone, and W. Dairman. Amino acid analysis with fluorescamine at the picomole level. Arch. Biochem. Biophys. 155:203-12.
