13 August 1918|
Rendcomb, Gloucestershire, England
19 November 2013
Cambridge, Cambridgeshire, England
|Alma mater||St John's College, Cambridge|
|Thesis||The metabolism of the amino acid lysine in the animal body (1943)|
|Doctoral advisor||Albert Neuberger|
Amino acid sequence of insulin
Frederick Sanger, OM, CH, CBE, FRS, FAA (; 13 August 1918 – 19 November 2013) was a British biochemist who won the Nobel Prize for Chemistry twice, one of only two people to have done so in the same category (the other is John Bardeen in Physics), the fourth person overall with two Nobel Prizes, and the third person overall with two Nobel Prizes in the sciences. In 1958, he was awarded a Nobel Prize in chemistry "for his work on the structure of proteins, especially that of insulin". In 1980, Walter Gilbert and Sanger shared half of the chemistry prize "for their contributions concerning the determination of base sequences in nucleic acids". The other half was awarded to Paul Berg "for his fundamental studies of the biochemistry of nucleic acids, with particular regard to recombinant DNA".
Early life and education
Frederick Sanger was born on 13 August 1918 in Rendcomb, a small village in Gloucestershire, England, the second son of Frederick Sanger, a general practitioner, and his wife, Cicely Sanger (née Crewdson). He was one of three children. His brother, Theodore, was only a year older, while his sister May (Mary) was five years younger. His father had worked as an Anglican medical missionary in China but returned to England because of ill health. He was 40 in 1916 when he married Cicely who was four years younger. Sanger's father converted to Quakerism soon after his two sons were born and brought up the children as Quakers. Sanger's mother was the daughter of a wealthy cotton manufacturer and had a Quaker background, but Cicely was not a Quaker.
When Sanger was around five years old the family moved to the small village of Tanworth-in-Arden in Warwickshire. The family were reasonably wealthy and employed a governess to teach the children. In 1927, at the age of nine, he was sent to the Downs School, a residential preparatory school run by Quakers near Malvern. His brother Theo was a year ahead of him at the same school. In 1932, at the age of 14, he was sent to the recently established Bryanston School in Dorset. This used the Dalton system and had a more liberal regime which Sanger much preferred. At the school he liked his teachers and particularly enjoyed scientific subjects.
He achieved good results in the School Certificate examinations and in 1936 moved as an undergraduate to St John's College, Cambridge to study natural sciences. His father had attended the same college. For Part I of his Tripos he took courses in physics, chemistry, biochemistry and mathematics but struggled with physics and mathematics. Many of the other students had studied more mathematics at school. In his second year he replaced physics with physiology. He took three years to obtain his Part I. For his Part II he studied biochemistry. It was a relatively new department founded by Gowland Hopkins with enthusiastic lecturers who included Malcolm Dixon, Joseph Needham and Ernest Baldwin.
Both his parents died from cancer during his first two years at Cambridge. His father was 60 and his mother was 58. As an undergraduate Sanger's beliefs were strongly influenced by his Quaker upbringing. He was a pacifist and a member of the Peace Pledge Union. It was through his involvement with the Cambridge Scientists' Anti-War Group that he met his future wife, Joan Howe, who was studying economics at Newnham College. They courted while he was studying for his Part II exams and married after he had graduated in December 1940. Under the Military Training Act 1939 he was provisionally registered as a conscientious objector, and again under the National Service (Armed Forces) Act 1939, before being granted unconditional exemption from military service by a tribunal. In the meantime he undertook training in social relief work at the Quaker centre, Spicelands, Devon and served briefly as a hospital orderly.
Sanger began studying for a PhD in October 1940 under N.W. "Bill" Pirie. His project was to investigate whether edible protein could be obtained from grass. After little more than a month Pirie left the department and Albert Neuberger became his adviser. Sanger changed his research project to study the metabolism of lysine and a more practical problem concerning the nitrogen of potatoes. His thesis had the title, "The metabolism of the amino acid lysine in the animal body". He was examined by Charles Harington and Albert Charles Chibnall and awarded his doctorate in 1943.
Neuberger moved to the National Institute for Medical Research in London, but Sanger stayed in Cambridge and in 1943 joined the group of Charles Chibnall, a protein chemist who had recently taken up the chair in the Department of Biochemistry. Chibnall had already done some work on the amino acid composition of bovine insulin and suggested that Sanger look at the amino groups in the protein. Insulin could be purchased from the pharmacy chain Boots and was one of the very few proteins that were available in a pure form. Up to this time Sanger had been funding himself. In Chibnall's group he was initially supported by the Medical Research Council and then from 1944 until 1951 by a Beit Memorial Fellowship for Medical Research.
Sanger's first triumph was to determine the complete amino acid sequence of the two polypeptide chains of bovine insulin, A and B, in 1952 and 1951, respectively. Prior to this it was widely assumed that proteins were somewhat amorphous. In determining these sequences, Sanger proved that proteins have a defined chemical composition. For this purpose he used the "Sanger Reagent", fluorodinitrobenzene (FDNB), to react with the exposed amino groups in the protein and in particular with the N-terminal amino group at one end of the polypeptide chain. He then partially hydrolysed the insulin into short peptides, either with hydrochloric acid or using an enzyme such as trypsin. The mixture of peptides was fractionated in two dimensions on a sheet of filter paper, first by electrophoresis in one dimension and then, perpendicular to that, by chromatography in the other. The different peptide fragments of insulin, detected with ninhydrin, moved to different positions on the paper, creating a distinct pattern that Sanger called "fingerprints". The peptide from the N-terminus could be recognised by the yellow colour imparted by the FDNB label and the identity of the labelled amino acid at the end of the peptide determined by complete acid hydrolysis and discovering which dinitrophenyl-amino acid was there. By repeating this type of procedure Sanger was able to determine the sequences of the many peptides generated using different methods for the initial partial hydrolysis. These could then be assembled into the longer sequences to deduce the complete structure of insulin. Finally, because the A and B chains are physiologically inactive without the three linking disulfide bonds (two interchain, one intrachain on A), Sanger and coworkers determined their assignments in 1955. Sanger's principal conclusion was that the two polypeptide chains of the protein insulin had precise amino acid sequences and, by extension, that every protein had a unique sequence. It was this achievement that earned him his first Nobel prize in Chemistry in 1958. This discovery was crucial for the later sequence hypothesis of Crick for developing ideas of how DNA codes for proteins.
From 1951 Sanger was a member of the external staff of the Medical Research Council and when they opened the Laboratory of Molecular Biology in 1962, he moved from his laboratories in the Biochemistry Department of the university to the top floor of the new building. He became head of the Protein Chemistry division. Soon after his move he started looking at the possibility of sequencing RNA molecules and began developing methods for separating ribonucleotide fragments generated with specific nucleases. One of the problems was to obtain a pure piece of RNA to sequence. In the course of this he discovered in 1964, with Kjeld Marcker, the formylmethionine tRNA which initiates protein synthesis in bacteria. He was beaten in the race to be the first to sequence a tRNA molecule by a group led by Robert Holley from Cornell University, who published the sequence of the 77 ribonucleotides of alanine tRNA from Saccharomyces cerevisiae in 1965. By 1967 Sanger's group had determined the nucleotide sequence of the 5S ribosomal RNA from Escherichia coli, a small RNA of 120 nucleotides.
He then turned to sequencing DNA, which would require an entirely different approach. He looked at different ways of using DNA polymerase I from E. coli to copy single stranded DNA. In 1975 together with Alan Coulson he published a sequencing procedure using DNA polymerase with radiolabelled nucleotides that he called the "Plus and Minus" technique. This involved two closely related methods that generated short oligonucleotides with defined 3' termini. These could be fractionated by electrophoresis on a polyacrylamide gel and visualised using autoradiography. The procedure could sequence up to 80 nucleotides in one go and was a big improvement on what had gone before, but was still very laborious. Nevertheless, his group were able to sequence most of the 5,386 nucleotides of the single-stranded bacteriophage φX174. This was the first fully sequenced DNA-based genome. To their surprise they discovered that the coding regions of some of the genes overlapped with one another.
In 1977 Sanger and colleagues introduced the "dideoxy" chain-termination method for sequencing DNA molecules, also known as the "Sanger method". This was a major breakthrough and allowed long stretches of DNA to be rapidly and accurately sequenced. It earned him his second Nobel prize in Chemistry in 1980, which he shared with Walter Gilbert and Paul Berg. The new method was used by Sanger and colleagues to sequence human mitochondrial DNA (16,569 base pairs) and bacteriophage λ (48,502 base pairs). The dideoxy method was eventually used to sequence the entire human genome.
As of 2013, he is the only person to have been awarded the Nobel Prize in Chemistry twice, and one of only four two-time Nobel laureates: The other three were Marie Curie (Physics, 1903 and Chemistry, 1911), Linus Pauling (Chemistry, 1954 and Peace, 1962) and John Bardeen (twice Physics, 1956 and 1972).
Marriage and family
Sanger married Margaret Joan Howe in 1940. They had three children — Robin, born in 1943, Peter born in 1946 and Sally Joan born in 1960. He said that his wife had "contributed more to his work than anyone else by providing a peaceful and happy home."
Sanger retired in 1983 to his home, "Far Leys", in Swaffham Bulbeck outside Cambridge and next to Sanger Wood.
In 1992, the Wellcome Trust and the Medical Research Council founded the Sanger Centre (now the Sanger Institute), named after him. The Institute is located on the Wellcome Trust Genome Campus near Hinxton, only a few miles from Sanger's home. He agreed to having the Centre named after him when asked by John Sulston, the founding director, but warned, "It had better be good." It was opened by Sanger in person on 4 October 1993, with a staff of fewer than 50 people, and went on to take a leading role in the sequencing of the human genome. The Institute now has over 900 people and is one of the world's largest genomic research centres.
Sanger said he found no evidence for a God so he became an agnostic. In an interview published in the Times newspaper in 2000 Sanger is quoted as saying: "My father was a committed Quaker and I was brought up as a Quaker, and for them truth is very important. I drifted away from those beliefs – one is obviously looking for truth, but one needs some evidence for it. Even if I wanted to believe in God I would find it very difficult. I would need to see proof."
He declined the offer of a knighthood, as he did not wish to be addressed as "Sir". He is quoted as saying, "A knighthood makes you different, doesn’t it, and I don’t want to be different." In 1986, he accepted the award of an Order of Merit, which can have only 24 living members.
In 2007 the British Biochemical Society was given a grant by the Wellcome Trust to catalogue and preserve the 35 laboratory notebooks in which Sanger recorded his research from 1944 to 1983. In reporting this matter, Science noted that Sanger, "the most self-effacing person you could hope to meet", was spending his time gardening at his Cambridgeshire home.
Sanger died in his sleep at Addenbrooke's Hospital in Cambridge on 19 November 2013. As noted in his obituary, he had described himself as "just a chap who messed about in a lab", and "academically not brilliant".
Awards and honours
The Wellcome Trust Sanger Institute (formerly the Sanger Centre) is named in his honour.
Selected publications by Frederick Sanger
- Neuberger, A.; Sanger, F. (1942), "The nitrogen of the potato", Biochemical Journal 36 (7–9): 662–671, .
- Neuberger, A.; Sanger, F. (1944), "The metabolism of lysine", Biochemical Journal 38 (1): 119–125, .
- Sanger, F. (1945), "The free amino groups of insulin", Biochemical Journal 39 (5): 507–515, .
- Sanger, F. (1947), "Oxidation of insulin by performic acid", Nature 160 (4061): 295, .
- Porter, R.R.; Sanger, F. (1948), "The free amino groups of haemoglobins", Biochemical Journal 42 (2): 287–294, .
- Sanger, F. (1949a), "Fractionation of oxidized insulin", Biochemical Journal 44 (1): 126–128, .
- Sanger, F. (1949b), "The terminal peptides of insulin", Biochemical Journal 45 (5): 563–574, .
- Sanger, F.; Tuppy, H. (1951a), "The amino-acid sequence in the phenylalanyl chain of insulin. 1. The identification of lower peptides from partial hydrolysates", Biochemical Journal 49 (4): 463–481, .
- Sanger, F.; Tuppy, H. (1951b), "The amino-acid sequence in the phenylalanyl chain of insulin. 2. The investigation of peptides from enzymic hydrolysates", Biochemical Journal 49 (4): 481–490, .
- Sanger, F.; Thompson, E.O.P. (1953a), "The amino-acid sequence in the glycyl chain of insulin. 1. The identification of lower peptides from partial hydrolysates", Biochemical Journal 53 (3): 353–366, .
- Sanger, F.; Thompson, E.O.P. (1953b), "The amino-acid sequence in the glycyl chain of insulin. 2. The investigation of peptides from enzymic hydrolysates", Biochemical Journal 53 (3): 366–374, .
- Sanger, F.; Thompson, E.O.P.; Kitai, R. (1955), "The amide groups of insulin", Biochemical Journal 59 (3): 509–518, .
- Ryle, A.P.; Sanger, F.; Smith, L.F.; Kitai, R. (1955), "The disulphide bonds of insulin", Biochemical Journal 60 (4): 541–556, .
- Brown, H.; Sanger, F.; Kitai, R. (1955), "The structure of pig and sheep insulins", Biochemical Journal 60 (4): 556–565, .
- Sanger, F. (1959), "Chemistry of Insulin: determination of the structure of insulin opens the way to greater understanding of life processes", Science 129 (3359): 1340–1344, .
- Milstein, C.; Sanger, F. (1961), "An amino acid sequence in the active centre of phosphoglucomutase", Biochemical Journal 79 (3): 456–469, .
- Marcker, K.; Sanger, F. (1964), "N-formyl-methionyl-S-RNA", Journal of Molecular Biology 8 (6): 835–840, .
- Sanger, F.; Brownlee, G.G.; Barrell, B.G. (1965), "A two-dimensional fractionation procedure for radioactive nucleotides", Journal of Molecular Biology 13 (2): 373–398, .
- Brownlee, G.G.; Sanger, F.; Barrell, B.G. (1967), "Nucleotide sequence of 5S-ribosomal RNA from Escherichia coli", Nature 215 (5102): 735–736, .
- Brownlee, G.G.; Sanger, F. (1967), "Nucleotide sequences from the low molecular weight ribosomal RNA of Escherichia coli", Journal of Molecular Biology 23 (3): 337–353, .
- Brownlee, G.G.; Sanger, F.; Barrell, B.G. (1968), "The sequence of 5S ribosomal ribonucleic acid", Journal of Molecular Biology 34 (3): 379–412, .
- Adams, J.M.; Jeppesen, P.G.; Sanger, F.; Barrell, B.G. (1969), "Nucleotide sequence from the coat protein cistron of R17 bacteriophage RNA", Nature 223 (5210): 1009–1014, .
- Barrell, B.G.; Sanger, F. (1969), "The sequence of phenylalanine tRNA from E. coli", FEBS Letters 3 (4): 275–278, .
- Jeppesen, P.G.; Barrell, B.G.; Sanger, F.; Coulson, A.R. (1972), "Nucleotide sequences of two fragments from the coat-protein cistron of bacteriophage R17 ribonucleic acid", Biochemical Journal 128 (5): 993–1006, .
- Sanger, F.; Donelson, J.E.; Coulson, A.R.; Kössel, H.; Fischer, D. (1973), "Use of DNA Polymerase I Primed by a Synthetic Oligonucleotide to Determine a Nucleotide Sequence in Phage f1 DNA", Proceedings of the National Academy of Sciences USA 70 (4): 1209–1213, .
- Sanger, F.; Coulson, A.R. (1975), "A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase", Journal of Molecular Biology 94 (3): 441–448, .
- Sanger, F.; Nicklen, S.; Coulson, A.R. (1977), "DNA sequencing with chain-terminating inhibitors", Proceedings of the National Academy of Sciences USA 74 (12): 5463–5467, . According to the Institute for Scientific Information (ISI) database, by October 2010 this paper had been cited over 64,000 times.
- Sanger, F.; Air, G.M.; Barrell, B.G.; Brown, N.L.; Coulson, A.R.; Fiddes, C.A.; Hutchinson, C.A.; Slocombe, P.M.; Smith, M. (1977), "Nucleotide sequence of bacteriophage φX174 DNA", Nature 265 (5596): 687–695, .
- Sanger, F.; Coulson, A.R. (1978), "The use of thin acrylamide gels for DNA sequencing", FEBS Letters 87 (1): 107–110, .
- Sanger, F.; Coulson, A.R.; Barrell, B.G.; Smith, A.J.; Roe, B.A. (1980), "Cloning in single-stranded bacteriophage as an aid to rapid DNA sequencing", Journal of Molecular Biology 143 (2): 161–178, .
- Anderson, S.; Bankier, A.T.; Barrell, B.G.; De Bruijn, M.H.; Coulson, A.R.; Drouin, J.; Eperon, I.C.; Nierlich, D.P.; Roe, B.A. (1981), "Sequence and organization of the human mitochondrial genome", Nature 290 (5806): 457–465, .
- Anderson, S.; De Bruijn, M.H.; Coulson, A.R.; Eperon, I.C.; Sanger, F.; Young, I.G. (1982), "Complete sequence of bovine mitochondrial DNA. Conserved features of the mammalian mitochondrial genome", Journal of Molecular Biology 156 (4): 683–717, .
- Sanger, F.; Coulson, A.R.; Hong, G.F.; Hill, D.F.; Petersen, G.B. (1982), "Nucleotide sequence of bacteriophage λ DNA", Journal of Molecular Biology 162 (4): 729–773, .
- Sanger, F. (1988), "Sequences, sequences, and sequences", Annual Review of Biochemistry 57: 1–28, .
- "SANGER, Frederick". Who's Who 2013, A & C Black, an imprint of Bloomsbury Publishing plc, 2013; online edn, Oxford University Press.(subscription required)
- "Seven days: 22–28 November 2013". Nature 503 (7477): 442. 2013.
- Allen, A. K.;
The Nobel Prize in Chemistry 1958: Frederick Sanger – biography, Nobelprize.org, retrieved 10 October 2010
The Nobel Prize in Chemistry 1980: Frederick Sanger – autobiography, Nobelprize.org, retrieved 10 October 2010
- A Life of Research on the Sequences of Proteins and Nucleic Acids: Fred Sanger in conversation with George Brownlee, Biochemical Society, Edina – Film & Sound Online, 9 October 1992. Subscription required. A 200 min interview divided into 44 segments. Notes give the content of each segment.
- Sanger, Frederick (1944). The metabolism of the amino acid lysine in the animal body (PhD thesis). University of Cambridge.
- Neuberger & Sanger 1942; Neuberger & Sanger 1944
- Chibnall, A. C. (1942). "Bakerian Lecture: Amino-Acid Analysis and the Structure of Proteins". Proceedings of the Royal Society B: Biological Sciences 131 (863): 136. Section on insulin starts on page 153.
- Sanger & Tuppy 1951a; Sanger & Tuppy 1951b; Sanger & Thompson 1953a; Sanger & Thompson 1953b
- Sanger, F. (1958), Nobel lecture: The chemistry of insulin, Nobelprize.org, retrieved 18 October 2010. Sanger's Nobel lecture was also published in Science: Sanger 1959
- Ryle et al. 1955.
- Stretton, A. O. (2002). "The first sequence. Fred Sanger and insulin". Genetics 162 (2): 527–532.
- The Nobel Prize in Chemistry 1958: Frederick Sanger, Nobelprize.org, retrieved 8 October 2010
- Marcker & Sanger 1964
- Holley, R. W.; Apgar, J.; Everett, G. A.; Madison, J. T.; Marquisee, M.; Merrill, S. H.; Penswick, J. R.; Zamir, A. (1965). "Structure of a Ribonucleic Acid". Science 147 (3664): 1462–1465.
- Brownlee, Sanger & Barrell 1967; Brownlee, Sanger & Barrell 1968
- Sanger et al. 1973
- Sanger & Coulson 1975
- Sanger, F. (1980), Nobel lecture: Determination of nucleotide sequences in DNA, Nobelprize.org, retrieved 18 October 2010
- Sanger et al. 1977
- Sanger, Nicklen & Coulson 1977
- The Nobel Prize in Chemistry 1980: Paul Berg, Walter Gilbert, Frederick Sanger, Nobelprize.org, retrieved 8 October 2010
- Anderson et al. 1981
- Sanger et al. 1982
- : Frederick Sanger, OMThe Telegraph (accessed 20 November 2013)
- Frederick Sanger, Wellcome Trust Sanger Institute, retrieved 12 October 2010
- Hargittai, István (April 1999), "Interview: Frederick Sanger", The Chemical Intelligencer (New York: Springer-Verlag) 4 (2): 6–11. This interview, which took place on 16 September 1997, was republished in Hargittai, István (2002), "Chapter 5: Frederick Sanger", Candid science II: conversations with famous biomedical scientists, London: Imperial College Press, pp. 73–83,
- Ahuja, Anjana (12 January 2000), "The double Nobel laureate who began the book of life",
- Bhattachjee, Yudhijit, ed. (2007), "Newsmakers: A Life in Science", Science 317: 879,
- "Double Nobel-winning 'father of genomics' Frederick Sanger dies". The Australian. 21 November 2013.
- "Frederick Sanger: Nobel Prize winner dies at 95". BBC.co.uk. 20 November 2013. Retrieved 20 November 2013.
- "Frederick Sanger: Unassuming British biochemist whose pivotal and far-reaching discoveries made him one of a handful of double Nobel prizewinners". The Times (London). 21 November 2013. p. 63.
- Frederick Sanger’s achievements cannot be overstated, The Conversation, 21 November 2013
- Finch, John (2008), A Nobel Fellow on every floor: a history of the Medical Research Council Laboratory of Molecular Biology, Cambridge: Medical Research Council, .
- Sanger, F.; Dowding, M. (1996), Selected Papers of Frederick Sanger: with commentaries, Singapore: World Scientific, .
- The Sanger Institute
- About the 1958 Nobel Prize
- About the 1980 Nobel Prize
- Fred Sanger 2001 Video Documentary by The Vega Science Trust
- National Portrait Gallery
- Frederick Sanger interviewed by Alan Macfarlane, 24 August 2007 (film), also available on Video on YouTube. Duration 57 minutes.
- Interviews with Nobel Prize–winning scientists: Dr Frederick Sanger, British Broadcasting Corporation, c. 1985. Interviewed by Lewis Wolpert. Duration 1 hour.
- Frederick Sanger archive collection - Wellcome Library finding aid for the digitised collection.