Christian B. Anfinsen the Journey Not the Attival Matters - Leonard Woolf, After Montaigne

Christian B. Anfinsen The journey not the aTTival matters - Leonard Woolf, after Montaigne

CHRISTIAN BOEHMER ANFINSEN entific director of the new institute died suddenly on May 14, 1995 at the and was vigorously recruiting scien age of 79. At the time he was Pro tists to staff a partially renovated Build fessor of Biophysical Chemistry at ing 3, built in l 938 largely as an ani the Johns Hopkins University, a po mal facility when NIH moved from sition he had assumed after his re Washington, D.C. Among those who tirement from the National Insti responded to Shannon's vision were tutes of Health (NIH) in 1981. In Julius Axelrod, Robert Berliner, Rob 1972 he shared, with Stanford ert Bowman, Bernard Brodie, Donald Moore and William H. Stein of Fredrickson, Edward Korn, Earl and Rockefeller University, the Nobel Thressa Stadtman, Daniel Steinberg, Prize for Chemistry. He had been and Sidney Udenfriend. They joined cited by the Swedish Royal Academy Leon Heppel, Bernard Horecker, of Sciences for his "studies on ribo Herman Kalckar, Arthur Kornberg nuclease, in particular the relation and others on the staff of the Experi ship between the amino acid se mental Biology and Medicine Insti quence and the biologically active tute. This ensemble of individuals, conformation." crowded into the very small building Chris, as he was known to even (but later dispersed on the NIH cam the most junior member of his pus after the Clinical Center opened laboratory, was the son of Norwe in 1953 ), was a crucial seed for the gian immigrants from Bergen who explosive growth of biomedical re had settled in western Pennsylvania. 1916- 1995 search at the NIH that followed. His father, a road construction en During the next five years, a whole gineer, later moved the family to the He returned to the Harvard Bio range of publications on plasma lipo Philadelphia area where Chris gradu logical Chemistry Department in protein metabolism emerged from ated from Swarthmore College in 1946. During the next decade he Chris' lab, as well as a continuing 1937 and went on to study organic worked on a variety of projects in stream ofpapers on protein structure. chemistry at the University of Penn what historically may now be seen as While this lipoprotein work was un sylvania. A fellowship from the the transition phase of biochemistry doubtedly related to the interests of his American-Scandinavian Foundation from the study of intermediary me new employer, it was never fondly re sent him to the Carlsberg Labora tabolism to that of molecular and membered by him, although the ap tory in Copenhagen in 1939 to study structural biology. He used mi proaches he and his colleagues devel enzyme-based micromethods. The cromethods ('Cartesian Divers') for oped were very important in the sub outbreak ofwar forced his return to the measuring metabolic processes and, sequent clarification of the genetic United States in 1940, but not before he with A.K. Solomon, pioneered the bases of the lipoprotein diseases. had the chance to see and understand use of stable and radioactive isotopes However, it was Fred Sanger's the horrors then gripping Europe. His for tracing metabolic pathways, in contemporaneous work on the amino unusually deep and active sense of so cluding the biosynthesis and degrada acid sequence of insulin that really cial responsibility certainly dated tion of proteins. A year (1947- 1948) excited Chris and provided the theme from that period, if not earlier. at the Medical Nobel Institute in for the remainder of his career. Chris entered the renowned Stockholm, in Hugo Theorell's labo Surely the technique could be made graduate program in Biological ratory, led to partial purification of to work to determine the primary Chemistry at Harvard Medical aconitase, with Jack Buchanan, and structure of an enzyme and perhaps School where he worked with further hints of his evolving interest ultimately to synthesize the protein. A. Baird Hastings on a problem in in proteins. The availability of a ready supply of retinal histochemistry. He was In 1950, to the astonishment of his bovine pancreatic ribonuclease awarded his Ph.D. degree in 1943; by Boston colleagues, Chris did the un (RNase) from the Armour Com 1944 he was working at Harvard in the thinkable and gave up his position as pany, a by-product of its protein malaria research project of Vannevar Associate Professor at Harvard and fractionation work, defined the en Bush's Office of Scientific Research moved to the pastures of Bethesda, zyme of choice. In 1954, the first and Development. Thirty years later Maryland to become Chief of the paper appeared from Chris's labo some of the observations Chris made Laboratory of Cellular Physiology in ratory on the structure, cross-link then were used in developing the the newly created National Heart In ages and terminal sequences of current methods of culturing ma stitute of the NIH. James Shannon RNase. Despite its apparent irrel laria parasites. had recently gone to Bethesda as sci- evance to heart dise,ise, this work structural biology volume 2 number 8 august 1995 621 © 1995 Nature Publishing Group http://www.nature.com/nsmb obituary •

was the beginning of fifteen years of perimental analyses of the refolding of in recent years work on chaperones concentrated effort on RNase. RNase and other proteins to lead to has been interpreted by some to con In 1954, Chris returned to the the full 'thermodynamic hypothesis'. tradict Chris' hypothesis. This con Carlsberg Laboratory on a one-year This hypothesis stated that 'the three fusion between thermodynamic and Rockefeller fellowship, to work with dimensional structure of a native pro kinetic analyses, common in many Kai Linderstr0m-Lang, using physical tein in its normal physiological milieu other areas of science, misses the fun chemistry to study RNase. He joined (solvent, pH, ionic strength, presence damental nature of the contribution. a remarkable group of scientists, in of other components such as metal In fact, Chris and his colleagues cluding Aase Hvidt, Martin Ottesen, ions or prosthetic groups, tempera Franco deLorenzo, David Givol and Bill Harrington and John Schellman ture, and others) is the one in which Sara Fuchs-were the first to identify who, over the next decades, contrib the Gibbs free energy of the whole sys and characterize a chaperone, the pro uted greatly to defining the properties tem is lowest; that is, that the native tein-disulphide isomerase. Although of globular proteins. Although, ironi conformation is determined by the to Chris participated actively in both ki cally, the main conclusion of the com tality of interatomic interactions and netic and equilibrium experiments bined efforts of this group on RNase hence by the amino acid sequence, in (including with this author), at heart that "the data further support the pos a given environment"'. he favoured a thermodynamic ap sibility that a considerable part of the This simple and elegant principle proach. He was more interested in enzyme structure may be superfluous has, over the last third-of-a-century, finding out what existed in nature or from the catalytic standpoint" did not become part of the fundamental para what one could synthesize, then the stand up to later work, the structural digm of molecular biology, as well as details of the process being studied. concepts that Chris added to his the basis of vast biotechnology efforts. From 1962 to 1963, Chris returned chemical and enzymological back The enormous skepticism at the time for a year to Harvard as Professor of ground were of major importance in among the scientific establishment to Biological Chemistry, but was re the next few years. both the experimental results and cruited back to the NIH in 1963 by Upon returning to the NIH it be their interpretation has now been J.E. Rall to head a newly created came apparent to him that the Moore largely forgotten. Indeed, to some the Laboratory of Chemical Biology in and Stein team at Rockefeller would concepts were considered--consistent the National Institute of Arthritis likely complete the amino acid se with J.B.S. Haldane's quip about the and Metabolic Diseases. At this quence of RNase first. Instead of an response to all new ideas-to have point a new model protein, staphy all-out effort to compete on the se been self-evident. On the other hand, lococcal nuclease, was added to his quence, he chose to focus on the dif ficult problem of defining the disulphide cross-links among the eight cysteine residues in RNase. In 1956, during work to develop revers ible cleavage techniques for the four cystine cross-links, he noted serendipitously that the disulphide bonds could reform with restoration of enzymatic activity, under certain conditions. It was at this moment that Chris' genius for getting to the es sence of research problems fully mani fested itself. He quickly realized that the unexpected reversibility of this process, which later included the demonstration of the restoration of secondary and tertiary structure as well as the disulphide bonds, im plied that the information for the folding of a protein is contained in the amino-acid sequence. The re sult was also of interest to him in that it implied that the synthesis of func tional proteins would only require the sequential polymerization of the ap propriate amino acids. The efforts over the next six years of Chris and his colleagues-especially Fred White, Michael Sela, Ed Haber, Charles Epstein and Robert Chris photographed in his office at the National Institutes of Health in the late 1970s Goldberger- supplied the detailed ex- with the typewriter (background) that he used to w rite all his notes and letters.

Linderstr0m-Lang's years following these studies and the treaty banning atmospheric nuclear caricature poster of award of the Nobel Prize, Chris con testing, in his opposition to the US Chris Anfinsen tributed significantly to the pu involvement in Vietnam, and in his exhibited at the Carlsberg Lab• rification of human interferon and human rights activism, especially oratory Christmas later to the study of highly thermo with regard to scientists in the USSR party, 1954 (Cour• stable enzymes. Indeed at the time of and in Latin America. For a number tesy of Carlsberg his death he was funded by the Na of years he chaired the human rights Foundation Picture tional Science Foundation for devel committee of the US National Acad Archives, Copen• oping thermostable enzymes for the emy of Sciences. hagen). remediation of environmental con Playing the piano or the viola and, tamination. most of all, sailing provided him with This summary of Chris' scientific limited escapes from science. In sail accomplishments would be incom ing, as in all things, he was always the plete without mention of several other optimist; his casualness in his sailboat aspects of his career. In 1959 he pub made these voyages only for the in lished the monograph The Molecular trepid. However, when crises occurred Basis of Evolution'. Appearing at the on the water, everyone knew that Chris time of the Darwin centennial, this would solve the problem, and he al book was important in initiating the ways did. current era of the use of macromo But above all of his professional ac lecular structure information in evo tivities was Chris' impeccable stan experimental program and over the lutionary analyses. It was highly in dards of personal behaviour, whether next decade it became the centre of fluential in shaping the perspective of he was your teacher, colleague, col his research interest. Its lack of many of us who entered science in the laborator, competitor, boss or friend. disulphide bonds and its great stabil following decade. His editorship, The roles melded completely into each ity made it a better experimental sys since 1957, of Advances in Protein other; all individuals, from the stock tem than RNase to study the process Chemistry, most recently with John clerk to the director of the Institutes, of protein folding by synthetically al Edsall, Fred Richards, and David were treated as fellow crew members tering individual amino-acid resi Eisenberg, is considered by many to in the quest to understand nature. dues. During this period Chris, and have been equally influential in defin Chris never looked back. Even the a very large number of young col ing the field of protein chemistry. Nobel lecture had to have new, leagues, used organic synthetic, Chris was also instrumental in unpublished data. Only death has fi chemical, enzymological, biophysi making the NIH much more like a nally stopped a research journey cal, immunological and genetic ap university than a typical government which began with his trip to proaches to study the properties of laboratory. He helped create The Copenhagen fifty-six years ago. this protein and its folding. A fold Foundation for Advanced Education ing mechanism incorporating'flick in the Sciences at the NIH and other Alan N. Schechter ering' secondary structure nucle teaching programs-modelJed after Laboratory of Chemical Biology, ation sites and subsequent conden the Swarthmore honors seminars NIDDK, National Institutes of Health, sation of tertiary structure was which made the NIH an unusually Bethesda, Maryland, USA 20892. evolved that is not much different excellent learning experience for the from today's concepts. By the mid many young scientists, especially phy Christian Boehmer Anfinsen, bio l 970's Chris' group, with the help of sicians, who came during his three chemist; born Monessen, Pennsylva a crystal structure determination by decades there. nia, 26 March 1916; B.A., Swarthmore F.A. Cotton and his associates, had Chris' laboratory was an interna College 1937; M.S., University of added 'staph' nuclease to the small tional mecca for scientists. As he said, Pennsylvania, 1939; Ph.D., Harvard number of globular proteins, in the best way to promote scientific ex University Medical School; professor, cluding hemoglobin, myoglobin, change among countries is to ex Harvard 1946-1950, 1962-1 963; labo lysozyme as well as RNase, which change scientists. Since his 1957 visit ratory chief, National Institutes of had been most thoroughly charac to Israel, at the invitation of Michael Health, l 950-l 962, I 963-I 98 I; profes terized. The goal of synthesis of nu Sela and Ephraim and Aaron sor, Weizmann Institute 1981-1982; clease and its analogues was partially Katchalski, Chris maintained a deep professor, Johns Hopkins, 1982-1995; achieved with 'semi-synthesis', by interest in that country and its science. Nobel Prize in Chemistry, 1972; mar condensation of fragments made by He frequently visited and served for riage to Florence Kenenger, l 941 -1978 the solid-phase method. Recombi almost four decades on the Board of (one son, two daughters); married nant DNA technologies ultimately Governors of the Weizmann Institute Libby Shulman Ely 1979; died made this goal fully practical. of Science in Rehovot. This deep in PikesviJle, Maryland, 14 May 1995. Along this reseach path Chris terest later extended to the Hebrew with Iku Kato, Meir Wilchek and language and the Jewish religion. 1. Anfinsen, C B. Principles that govern the to political issues folding of protein chains. (Nobel Lecture) Pedro Cuatrecasas-was also key in es Chris' sensitivity Science 181, 223-230 (1973). tablishing affinity chromatography as was manifest, among other activities, 2. Anfinsen, C B. The Molecular Basis of a major tool for biochemistry. In the in his work leading up to the 1963 Evolution (John Wiley, New York; 1959). 623 structural biology volume 2 number 8 august 1995