Double Nobel prize winner: Frederick Sanger – the father of genomics

This paper aims to outline briefly the main stages of Frederick Sanger’s scientific activity – the only person to have won two Nobel Prizes in Chemistry (1958, 1980). His work on the structure of proteins, especially that of insulin, and the determination of base sequences in nucleic acids made an immense impact on the development of biochemistry and especially on the development of a new scientific field – molecular biology. His methods for determining the primary structure of proteins and nucleic acids helped biochemists and molecular biologists to determine the structure of many proteins and nucleic acids and laid the basis for genetic engineering.

T he Nobel Prize is considered the highest honor inthescientificcommunity.Reflecting the highest achievements one can attain in socie ty, it has become more than just a prize -it provides worldwide recognition, respect, and prestige. It is a great honor to deliver a Nobel lecture, and even a greater honor to deliver it twice. Between 1901 and 2020, 934 Laureates and 28 organizations have been awarded the Nobel Prize. Four scientists obtained a Nobel Prize twice -Marie SkłodowskaCurie(Physics 1903,Chemistry1911), LinusPauling (Chemistry1954,Peace1962),John Bardeen(Physics 1956,Physics1972),andFrederick Sanger(Chemistry1958,Chemistry1980) [1].Frede rickSangerandLinusPauling'sscientificcreativity was directly related to the central focus of generating newLifeScienceknowledge [2].Thispaperwede-votetoFrederickSangerandhisscientificlegacy.
frederick sanger was born on August 13, 1918,inthevillageofRendcomb(Gloucestershire, England).Hewasthemiddlechildinthefamilyof Frederick Sanger, a country medical doctor, and his wifeCicelySanger(néeCrewdson),thedaughterof a wealthy cotton manufacturer. Cicely and Frederick Sanger had three children: Theodore was born in 1917, Frederick -in 1918, andMary -in 1923. All children were brought up as Quakers as their father was a devoted religious man active in theSocietyofFriends [5].Thechildrenweretaught such values as truth and hard work that defined Frederick's personality. Even when he no longer followed Quaker belief, these personal traits remained. When Frederick was 5 years old, his family moved in Tanworth the University of Cambridge as an undergraduate , I had to decide which three scientific subjects I should take. I had chosen two-and-a-half subjects and was looking through the list of "half" subjects when I noticed one I had not heard of before: "Biochemistry, supervisor Ernest Baldwin". The idea that biology could be explained in terms of chemistry seemed an exciting one, and this was amply confirmed when I met the enthusiastic Dr. Baldwin" [7].
Ernest Baldwin was a member of the Biochemistry Department of the University of Cambridge established by an English biochemist Frederick Gowland Hopkins -the Nobel Prize winner in Physiology or Medicine 1929 for the discovery ofvitamins [8].Hopkinshadtheenthusiasticsupport of his younger colleagues. Joseph and Dorothy Needhams dubbed him the Fundator et Primus Ab-basofbiochemistryinEngland [9].In1961duringa Symposium on Biochemistry and Nutrition to celeb-ratethecentenaryofthebirthofFrederickGowland Hopkins,JosephNeedhamstated:"our meeting today symbolizes the feeling of discipleship which we all have for Frederick Gowland Hopkins, essentially the founder of modern biochemistry in our country" [10].WhenFrederickGowlandHopkinsretired,Joseph Needham moved out of biochemistry to focus on his other scope of interest -Chinese civilization and science. The Department of Biochemistry headed byHopkinsbecameaninteresting andex-citingplaceforFrederickSanger.Asheexpected, it was here where he had a chance to acquire the necessary fundamental knowledge on living matter forsolvingproblemsinthefieldofmedicine.
During his second year at the University, Frede rick dropped physics and chose to study physio logy instead, keeping his studies in chemistry, biochemistry , and math, which he was not happy with.Helikedbiochemistrymostanditbecamethe subjectforPartIIoftheTripos [5,6].
In 1936, Frederick Sanger joined the Cambridge Scientists' AntiWar Group -a leftwing pacifistgroupfoundedin1932tocampaignagainst militarism.JosephNeedham-oneofhislecturers inbiochemistry-wasalsoamemberoftheGroup. Sanger's religious background to a large extent shaped his pacifist views. Here he met his future wife Joan Margaret Howe. She was an economics student at Newnham College. They married in 1940 and remained married until her death in 2012. They hadthreechildren-Robin(1943),Peter(1946),and SallyJoan(1960).FrederickSangerascribedhiswife Frederick Sanger [3] Mary, Frederick and Theodor Sanger [4] science and having a good rapport with his biology master Frazer Hoyland and chemistry master Henry Geoffrey Ordish, he was involved in biologyandchemistry clubs [6].Doingwellatschool, he passed the School Certificate exams on seven subjectsthatquali fiedhimforentrytoCambridge University.In1936,hewasacceptedbySt.John's College. Frede rick's parents had been hoping their son would follow his father's footsteps by studying medicine.However,ayoungstudentabandonedthe idea as he believed that science would provide him a more suitable lifestyle and would not be so timeconsuming.
In Cambridge for Part I of the Natural Scien ces Tripos, Frederick Sanger took chemistry, physics , andmathandbiochemistryashalfsubjects [6].Explaining his choice of biochemistry, terra incognita forhimatthattime,Sangerwrote:"When I arrived at akeyroleinhissuccessfulcareer,saying:"Although not a scientist herself she has contributed more to my work than anyone else by providing a peaceful and happy home" [11].
In 1939, he graduated in biochemistry from St.John'sCollege,Cambridge.EarningaFirston hisbiochemistryexam,hewassurprised.Beinga modestperson,Sangerwrote:"I was not academically brilliant. I never won scholarships and would probably not have been able to attend Cambridge University if my parents had not been fairly rich…" [7].DuringWorldWarII,atribunalgrantedSanger conscientious objector status and he became an orderly at a military hospital near Bristol. Frederick went to Cambridge to become a research student. As farashedidnotneeduniversityfinancialsupport,he gainedadmission [5].HestartedhisPhDinOctober 1940 under the supervision of Norman Wingate (Bill) Pirie.Heaimedtoinvestigatewhetheredibleprotein couldbeobtainedfromgrass.However,Pirieleftthe department, and Albert Neuberger was assigned as Frederick's new mentor. Sanger changed his thesistitleandworkedonthetopic"The metabolism of the amino acid lysine in the animal body" [12]. Describing thebeginningofhisscientificjourney, Sangerstated:"I regard Albert as my main teacher. The most important thing he taught me, both by instruction and by example, was how to do research. I shall always be grateful for his kindness and patience. He also had an extremely wide knowled ge of biochemistry, which I admired and used but could never emulate" [7].
Frederick Sanger received his PhD in 1943. The emphasis on chemistry in Sanger's thesis was to be themainstayofhisfuturescientificproject.Afterreceiving his doctorate, Sanger joined a research group led by albert Charles Chibnall,whoreplacedHopkins as a professor of biochemistry at Cambridge. A research group led by him at the time was studying the chemistry of proteins including the structure of insulin.
Working with Chibnall, Sanger became engaged in the identification of free amino groups in insulin. That time was particularly successful for protein chemistry. New methods have been developed for the fractionation of biopolymers, and therewasarealopportunitytodeterminetheexact chemical structure of these fundamental components of living matter. Sanger's interest in chromatography methods developed by the British biochemists archer martin and Richard Synge, the 1952 Nobel laureatesinchemistry [13],promptedhimtobegin work on determining protein structure.
Using insulin as a model for study, Sanger developed a new method for analyzing the structure of proteins and showed that the insulin molecule is composed of two peptide chains referred to as the A chain and B chain. They are linked together bytwodisulfidebonds,andinmostspecies,theA chain consists of 21 amino acids and the B chain of 30aminoacids.Benefitingfromthemethodofpaperchromatography,Sangeridentifiedunmodified aminoacids [14,15].Ittookhimyearstodefinitively identify 51 amino acids in the molecule of this proteinhormone [16].
F. Sanger worked out the sequence of 30-residue-long B chain together with the Austrian scientist Hans Tuppy and 21-residue-long A chain with Ted Thompson,aPhDstudentfromAustralia [1721].
In 1955, Sanger completed his investigation on the insulin sequence, and his work later became the basis for the production of synthetic insulin and other hormones. From 1944 to 1951, Frederick Sanger held a Beit Memorial Fellowship for Medical Research and since 1951 he has been a member of the ExternalStaffoftheMedicalResearchCouncil.
In 1958, Frederick Sanger received the Nobel PrizeinChemistry"for his work on the structure of proteins, especially that of insulin" [3]. In his NobelLecture"TheChemistryofInsulin",Sanger stressed:"The determination of the structure of insulin clearly opens up the way to similar studies on other proteins and already such studies are going on in a number of laboratories. These studies are aimed at determining the exact chemical structure of the many proteins that go to make up living matter and hence at understanding how these proteins perform their specific functions on which the processes of life depend. one may also hope that studies on proteins may reveal changes that take place in disease and that our efforts may be of more practical use to humanity" [22]. Frederick Sanger's discovery made it possibleto"lookinside"theproteinmoleculeandthus opened a new era in the development of modern biochemistry -protein chemistry.
Years after Nobel Prize award Sanger called "leanyears".ButmovingtothenewMRCLaboratory of Molecular Biology and joining forces with the group headed by Max Perutz marked a new pe-riodinFrederick'sscientificlife.Oneofthreedivisions was allotted to Sanger's group. Nucleic acids that were not of much interest before came to the fore of Sanger's scientific research. According to Sanger,"with people like Francis Crick around, it was difficult to ignore nucleic acids or to fail to realize the importance of sequencing them. An even more seminal influence was John Smith, who was the nucleic acid expert in the new laboratory and who was extremely helpful to me, so that I could turn to him for advice in this new field" [7].
Although nucleic acids as experimental material was new for Frederick, his interest in the problem of determining the primary sequence of biomole cules -sequencing -remained unchanged. Turning attention to the nucleic acids, RNA and DNA, Frede rick Sanger developed methods for determining small sequences in RNA. And later, hedevelopedthe"dideoxy" technique for DNA sequencing [25].Thiswasarelativelyrapidmethod and was used to determine the DNA sequence of the bacteriophageφX174of5375nucleotidesin1977, ofhumanmitochrondrialDNA (16,338nucleotides) and of bacteriophage λ (48,500 nucleotides) [11]. Thismethodisalsoreferredas"Sanger's method of DNA sequencing ".Sangerstated:"I suppose the dideoxy method can be regarded as the climax of my research career and the fulfilment of an ambition that had gradually been forming as I became more and more involved in sequencing" [7].sanger's sequencing was effectively adopted in the Human Genome Project (2000) which decoded the threebillion-letters human genetic code.
Sanger has repeatedly emphasized that this work owed much to his highly qualified collaborators. He had a high regard for B. G. Barrell, a. r. Coulson, and G. G. Brownlee, as well as for the students and postdoctoral fellows who had worked in the laboratory for several years.
In 1980, Frederick Sanger earned his second Nobel Prize in Chemistry jointly with Paul Berg and Walter Gilbert.OnehalfwasawardedtoPaulBerg "for his fundamental studies of the biochemistry of nucleic acids, with particular regard to recombinant-DNa",theotherhalf-jointlytoWalterGil-bertandFrederickSanger"for their contributions concerning the determination of base sequences Frederick Sanger at his Nobel Prize celebration (1958) [23] Frederick Sanger (left)

with Sydney Brenner (center) and Max Perutz (right) at a social event at the MRC Laboratory of Molecular Biology in Cambridge [24]
Frederick Sanger -the father of genomics looking at an autoradiogram of DNA sequence. MRC Laboratory of Molecular Biology [24] in nucleic acids" [26]. An American physicist, biochemist ,andmolecularbiologistWalterGilbert and his student Allan Maxam independently devised anewtechniqueforsequencingDNAin1977 [27]. This amazing coincidence indicates that ideas are in theair [28,29] [30] The Wellcome Trust Sanger Institute [32] as one of the greatest scientists of any generation, Sanger remained a very modest person throughout hislife.Heturneddowntheofferofanhonorarytitle of knighthood as he was against the idea of being addressedas"Sir".Hestatedthataknighthoodmade peopledifferent,andhedidnotwanttobedifferent [12]. Sanger's scientific discoveries have had a huge impact on the development of biochemistry and especially on the development of a new scientific field -molecular biology. His methods for determining the primary structure of proteins and nucleic acids helped biochemists and molecular biologists to determine the structure of many proteins and nucleic acids and laid the basis for genetic engineering. У статті представлено короткий огляд основних етапів життєвої та наукової діяльності Фредеріка Сенгера -єдиної людини, яка двічі отримала Нобелівську премію з хімії (1958,1980). Його роботи з вивчення структури протеїнів, особливо інсуліну, та встановлення послідовностей основ у нуклеїнових кислотах справиливеличезнийвпливнарозвитокбіохімії і, зокрема, на розвиток нової наукової галузімолекулярної біології. Його методи визначення первинноїструктурибіомакромолекулдопомоглибіохімікамімолекулярнимбіологамвстановитиструктурубагатьохпротеїнівінуклеїнових кислотізаклалипідвалинигенноїінженерії.