CHARLES SAMUEL HANES 21 May 1903—6 July 1990

( X o a / u J .

CHARLES SAMUEL HANES

21 May 1903—6 July 1990

Elected F.R.S. 1942

BY J.T.-F. WONG AND W. THOMPSON

CHARLES SAMUEL HANES was bom and raised in Toronto and received his elementary and secondary education in local schools. An enthusiastic and omnivorous student, he completed high school at the age of 16. Before entering the University of Toronto in 1920 he held several jobs including an assistantship in an instrument-making shop attached to the Weather Bureau, which experience he was to find invaluable. His duties there included flying a one metre weather balloon each morning and this he continued to do after his admission to the University of Toronto.

At University he studied all the sciences, eventually with an emphasis on biology and chemistry, and he graduated with high honours in 1925. The award of a coveted Overseas Scholarship of the Exhibition of 1851 took him to Cambridge which, with a few intermptions, was to be his base of operations for more than a quarter of a century. He was to receive a Ph.D. degree from Cambridge University in 1929 and an Sc.D. degree in 1953. He was elected to the Fellowship of the Royal Society in 1942 and to the Fellowship of the Royal Society of Canada in 1956.

Hanes arrived in Cambridge in 1925 when biological sciences in general were in a state of rapid development, and there he came into direct contact with, and was greatly influenced by, many of the leading scientists of the day, including F.F. Blackman, G.E. Briggs and F.G. Hopkins. But most influential of all was Theodora B. Auret, a research student in botany and plant pathology from South Africa, who took her Ph.D. (Cantab.) in 1932. She and Hanes were married in 1931 and together they forged a deeply affectionate, sustaining and companionable partnership over a life-time.

Hanes always remembered with satisfaction and pride his good fortune in being invited to become an Associate of the Ray Club, founded in 1937 in memory of the great 17th century naturalist, John Ray, F.R.S. The 12 Senior Members of the Club, who included at the time J.J. Thomson, E. Rutherford, H.F. Newall and F.G. Hopkins, met twice a year for dinner and conversation with the junior associates. Hanes was not only inspired by the calibre of these great scientists but also impressed by their kindness, humility and humour.

It was F.F. Blackman, reader in plant pathology and discoverer of the ‘dark reaction’ of photosynthesis, who suggested to Hanes in 1925 that he explore the mechanisms involved in the interconversions of starch and soluble sugars in plant tissues. So began his investigations of starch, the amylases, and later the plant phosphorylases. These studies were

149 © 1993 The Royal Society

150 Biographical Memoirs

continued at the Low Temperature Station for Research in Biochemistry and Physics (LTRS) where he came under the influence of W.B. Hardy, then director of Food Investigations. In 1931 Hanes returned to Toronto, hoping to find a teaching post, but times were hard. However, H.B. Speakman, director of the newly formed Ontario Research Foundation, offered him a fellowship and he worked for three years at the Foundation with a little part-time teaching at the University of Toronto. He returned to Cambridge in 1934 when he was offered an appointment on the staff of the LTRS.

THE VALUE OF ENZYMES IN THE STUDY OF POLYSACCHARIDES

In 1925, views on the structure of polysaccharides were confusing. Even the traditional ‘saccharide chain’ concept of Emil Fischer was questioned by certain prominent chemists who proposed that polysaccharide molecules were in reality very small ‘elementary units’, endowed with a mysterious tendency to clump together into colloidal aggregates. Indeed, not until the early 1930s was hard chemical evidence obtained by W.N. Haworth, E.L. Hirst and others to establish the basic chain structures of starch, glycogen and other polysaccharides. Meanwhile Hanes found that the (3-amylase of barley attacks only the free ends of a chain structure, liberating successive maltose fragments, whereas the a-amylase of malt attacks more deeply within the starch molecule. He recognized the value of enzymes as selective tools for probing starch structures and gradually persuaded Haworth and his colleagues at Birmingham of the validity of this approach. Invited by the editors of New Phytologist to review the subject, he received warm encouragement from Haworth. Hanes always regarded the writing of this review article, ‘The action of amylases in relation to the structure of starch and its metabolism in the plant’, a most salutary experience, causing him to organize his views toward a sizeable scientific problem and giving him new perspectives and improved analytical approaches. In the review, published in 1937, he not only provided an important early illustration of the usefulness of enzymes as probes of macromolecular structures but also suggested, on the basis of quantitative studies of the iodine-colouring property of starch and starch dextrins, that the saccharide chain of starch can exist in a helical conformation (see figure 1), which is essential for the iodine colouration; subsequent support for a helical arrangement has come from a variety of experimental approaches. This first

v? y° U° u ° l)°f °J tl

F ig u r e 1. Starch helix proposed by Hanes (Reproduced horn New Phytologist, 36, 101-189 (1937).)

Charles Samuel Hanes 151

proposal of a helical conformation for a biological macromolecule, made in 1937, represents a conceptual landmark of modern biochemistry. It is now well known that the tendency to helix formation extends also to proteins and nucleic acids.

Th e in f l u e n c e o f g .e . Br ig g s

In the Cambridge Botany School, Hanes had daily contact with G.E. Briggs for whom he developed a life-long affection and admiration. Briggs had just published with J.B.S. Haldane the steady-state solution of enzyme kinetics, and an outcome of this association with Briggs was the Hanes plot of S/v versus S for a hyperbolic rate curve which he used in a study of the action of amylase on starch. His interest in enzyme kinetics led decades later to the formulation of kinetic criteria for the analysis of complex enzyme mechanisms, the delineation of the half-branching, half-linear mechanism of liver alcohol dehydrogenase, and the evaluation of all ten rate constants for this mechanism from steady-state measurements.

Hanes is probably best known for his discovery and initial characterization of the plant phosphorylases, which he reported upon somewhat hurriedly before he left Cambridge for wartime duties. J. Bodnar and later B. Tanko, in Budapest, had shown that esterification of inorganic phosphate and the accumulation of hexose-phosphates occurred in suspensions of pea flour; the source of the hexose was unknown. Hanes listened with fascination to a lecture in Cambridge by J.K. Parnas describing the observations he and T. Baranowski had made on the breakdown of glycogen in dialysed muscle juice in the presence of phosphate. It was then a very simple matter, as Hanes retold it years later, for him to think of starch as the probable source of the hexose-phosphates in the pea flour suspensions. He proceeded then to demonstrate that dried peas, and other plant tissues, contain an extractible enzyme which catalyses the reversible conversion of starch into glucose-1-phosphate, or Cori ester.

PRODUCTION OF SYNTHETIC STARCH

Hanes and his assistant, E.T. Whitmore, laboured day and night to produce a large specimen of synthetic starch (about 110 g) by allowing purified potato phosphorylase to act upon glucose-1-phosphate. The bulk of this specimen was sent to Birmingham for examination by the methylation method, which showed it to contain (1—>4)-linked a-D-glucose residues having unmistakably the basic chain structure of starch. The material displayed the typical X-ray diffraction pattern of precipitated starch. Hanes also found it to be degraded by a- and (3-amylases in the same manner as amylose. This specimen of synthetic starch constituted the first example of a macromolecule, synthesized by the action of an enzyme in vitro, that could be identified with a natural product. Hanes recalled how close his precious specimen came to being eaten by Lord Woolton, the wartime Minister of Food in Britain. Franklin Kidd, then head of the LTRS, was entertaining the Minister at a lunch comprising delicacies such as grass soup, frozen whale meat and various dehydrated foods developed at the LTRS, and he pleaded with Hanes to donate the synthetic starch to be baked into biscuits to go with the soup!

The LTRS was part of the research organization created by Hardy during World War I to bring science to bear on the problems of food supplies in wartime. He attracted scientists

152 Biographical Memoirs

competent in all the basic areas relevant to the practical storage, transport and processing of foodstuffs, yet encouraged them to embark upon long-range research in their respective fields. Beginning in 1939, more and more of the basic research was shelved and, under the leadership of Franklin Kidd, attention was focused sharply upon wartime problems. Hanes shifted his efforts to practical matters, and served from 1941-1944 as scientific member of the British Food Mission in North America. Dividing his time between Washington, Ottawa, London and Cambridge, Hanes worked incessantly to resolve a kaleidoscopic sequence of technical problems connected with the production and handling of wartime food supplies. In 1943 he was appointed director of Food Investigation, and remained in this administrative post until 1947, when he was appointed Reader in Biochemistry at Cambridge and Professorial Fellow of Downing College.

HANES RETURNS TO THE LABORATORY

He returned happily to the laboratory - after the break of seven years - and enjoyed close associations with both the Botany School and the Department of Biochemistry: G.E. Briggs and A.C. Chibnall had collaborated in proposing his joint appointment to both departments. With F.A. Isherwood (of the LTRS) he explored many aspects of the then-new tool, paper chromatography; they developed the chromatographic separation of phosphoric esters, devising a spray that is still used to detect these esters. Hanes, F.J.R. Hird and Isherwood discovered several transpeptidases, including y glutamyl transpeptidase. Together with y glutamyl cyclotransferase, later discovered by G.E. Connell and Hanes, this enzyme plays a key role in the y glutamyl cycle of reactions involved in the transport of amino acids in tissues. It has also proved to be a valuable marker enzyme for liver carcinogenesis.

During his long career Hanes served on a variety of committees, boards and study groups on both sides of the Atlantic. For example, in wartime in the U.S.A. he became Chairman of the Committee on Dehydrated Foods of the Combined Food Board. In Britain he served on the Sub-Committee of the Scientific Advisory Committee on Utilization of National Resources, the Research Advisory Committee of the Forestry Commission and the Council of the Food Manufacturers Research Association; and in Canada, such Committees as the Committee of the National Research Council and the Research Advisory Committee of the Ontario Cancer Institute. For several years he was Chairman of the Canadian National Committee for Biochemistry. He recalled with great satisfaction the work he did with A.C. Chibnall when, for several years after the War, the two formed an informal Working Party and spent one week each year visiting in turn the Research Establishments of the Agricultural Research Council in Britain to enquire and support the work of the small groups of plant biochemists scattered throughout the country. In both Cambridge and Toronto he served on university boards, councils, degree committees, etc. Of particular importance was his contribution to the reorganizing of the School of Graduate Studies of the University of Toronto, under the Chairmanship of Bora Laskin who was to become Chief Justice of Canada.

The several lines of investigation Hanes had pursued at Cambridge were continued and expanded when he returned to Toronto in 1951. Refinement of paper chromatography of amino acids and peptides permitted the characterization and quantification of the diverse

Charles Samuel Hanes 153

products of transpeptidation. Kinetic investigations of sucrose phosphorylase and liver alcohol dehydrogenase were done and elastase was used to elucidate the structure of elastin. During this period G.E. Connell, G.H. Dixon, R.G. Donovan, C.K. Harris, A.T. Matheson, M.A. Moscarello, E.H.M. Wade, K.A. Walsh, T.E. Webb and J.T. Wong received their graduate training in his always-meticulous laboratory.

Hanes’s fine personal qualities, his open friendliness and generosity won him a legion of friends. He made a deep and lasting impression on those who had worked with him and his advice on all manner of problems was constantly sought. Hanes always had an aversion to ‘big’ science. In his view science in the universities was best pursued by small groups comprising a professor, a technician, two or three graduate students and the occasional postdoctoral fellow, a precept that he always followed, sustained by modest research grants. It was his opinion that the post -Sputnikdecade in North America, with the prospect of obtaining large research grants, served to convert several otherwise good scientists into ‘mere promoters and science managers’, whereas the modest organization that he favoured permitted the professor to remain an active scientist, involved in both teaching and research. He was an outstanding example of this approach.

Hanes retired from the headship of the Department of Biochemistry in Toronto in 1965, and became Professor Emeritus in 1968. He and his wife, Theo, for the most part continued to live quietly in Toronto sharing simple pleasures. These included a deep interest in their family, in gardening, music and in Spanish language and poetry. Over the years he remained in touch with his junior colleagues and sustained a lively interest in science. In the later years his wife’s ill health caused him great concern and he devoted much time to her care. He is survived by his wife, daughter and two grandchildren.

The portrait photograph shows Hanes at the University of Toronto. It was taken in 1979 by Mr M. Sawyer.

B ib l io g r a p h y

(1)(2)(3)

(4)

(5)

(6)

(7)(8 )

(9)

( 10)

( 11)( 12)

1927 Formation of resin in seedling confifers. Linnean Soc.1929 Method for determination of sugars. Biochemical 23, 99.1931 (With J. Barker) Effects of cyanide upon the respiration and sugar content of the

potato. Proc. R. Soc. Lond. B 108, 95.1932 Effect of starch concentration upon velocity of hydrolysis by the amylase of germinated

barley. Biochemical J. 26, 1406.1935 Reversible inhibition of beta-amylase by ascorbic acid and related compounds.

Biochemical J. 29, 2588.Action of the two amylases of barley. Can. J. Research B 13, 185.

1936 Determination of starch in plant tissues. Biochemical J. 30, 168.1937 The action of amylase in relation to the structure of starch and its metabolism in the

plant. (Monograph appearing in two parts) The New Phytologists 36, 102 & 189.1938 (With M. CATTLE) Starch-iodine coloration as an index of differential degradation by the

amylases. Proc. R. Soc. Lond. B 125, 387.1940 Breakdown and synthesis of starch by an enzyme system from pea seeds. Proc. R. Soc.

Lond. B 128, 421.Enzymic synthesis of starch from glucose-1-phosphate. Nature 145, 348.Reversible formation of starch from glucose-1-phosphate, catalysed by potato phosphorylase. Proc. R. Soc. Lond. B 129, 174.

154 Biographical Memoirs

(13) 1942

(14) 1946

(15) 1949

(16) 1950

(17) 1952

(18)

(19) 1953(20) 1954

(21) 1956

(22) 1959

(23) 1955

(24) 1961

(25)

(26)

(27)

(28)

(29)

(30)

(31) 1962

(32) 1963

(33) 1964

(34) 1965

(35)

(With E.J. MASKELL) Influence of hydrogen-ion concentration upon the equilibrium state in phosphorylase systems. Biochemical J. 36, 76.Report of the Food Investigation Board for the years 1940-1945. H.M. Stationery Office, London.(With F.A. ISHERWOOD) Separation of phosphoric esters on the filter paper chromatogram. Nature164, 1107.(With F. HlRD & F.A. ISHERWOOD) Synthesis of peptides in enzymic reactions involving glutathione. Nature 166, 288.(With F. HlRD & F.A. ISHERWOOD) Transpeptidation reactions involving y-glutamyl peptides and a-aminoacyl peptides. Biochemical J. 51, 25.(With G.E. CONNELL & G.H. Dixon) Transpeptideation and transamidation reactions. Phosphorus metabolism II, 95.Formation of peptides in enzymic reactions. Brit. Med. Bull. 9, 131.(With G.E. CONNELL & G.H. Dixon) Glutathione in relation to transpeptidation reactions. Glutathione Symposium. Academic Press, p. 145.(With G.E. CONNELL) Enzymic formation of pyrrolidone carboxylic aid from y-glutamyl peptides. Nature 177, 377.(With A.T. MATHESON) The chemical nature of intracellular peptidases. Bioch. Biophys. Acta 33, 292.Papers 23-29 are separate parts of a series devoted to the development of quantitative methods for the analysis of aminoacids in complex mixtures, e.g. in hydrolysates of protein; this work was done before the introduction of column chromatrography for the purpose.(With G.E. CONNELL & G.H. DIXON) Quantitative chromatographic methods for the study of transpeptidation reactions. Can. J. Bioch. Phys. 33, 416.Part 2. An approach to paper chromatography of improved resolving power and reproducibility. Can. J. Bioch. Phys. 33, 119.(With E.H.M. WADE & A.T. MATHESON) Part 3. Factors controlling patterns of separation of amino acids on paper chromatograms. Can. J. Bioch. Phys. 39, 141 (With C.K. HARRIS, M.A. MOSCARELLO & E. Tigane) Part 4. Stabilized chromatographic systems of high resolving power for amino acids. Can. J. Bioch. Phys. 39, 163.(With A.T. MATHESON & E. TIGANE) Part 5. An improved ninhydrin-hydrindantin reagent. Can J. Bioch. Phys. 39, 417.(With E. TIGANE, E.H.M. Wade & J.T-F. WONG) Part 5. Improved procedure for determination of amino acids. Can. J. Bioch. Phys. 39, 427.(With C.K. Harris & E. TIGANE) Part 7. Isolation of amino acids from serum and other fluids. Can. J. Bioch. Phys. 39, 439.(With M.A. MOSCARELLO & B.G. LANE) Chromatographic systems of high resolving power for nucleotides. Can. J. Bioch. Phys. 39, 1755.(With J. TZE-FEI WONG) Kinetic formulations for enzymic reactions involving two substrates. Can. J. Bioch. Phys. 40, 763.(With A.T. MATHESON & S. BJERRE) A conjugated form of amino-peptidase from autolysed extracts of kidney tissue. Can. J. Bioch. Phys. 41, 1741.(With J. TZE-FEI WONG) Isotropic exchange at equilibrium as a criterion of enzymatic mechanisms. Nature 203, 492.(With R.A. A n w a r & R.G. Do n o v a n ) Comparison of elastins from various sources. Proc. Third Can. Conference on Rheumatic Diseases.(With R.G. DONOVAN & R.A. A n w a r ) Picture of the breakdown of elastin under the action of elastase. Proc. Third Can. conference on Rheumatic Diseases.

Charles Samuel Hanes 155

(36) 1969

(37) 1972

(38)

(39)

(40) 1973

(41)

(42) 1978

(With J.T-F. WONG) A novel criterion for enzymic mechanisms involving three substrates. Archs. Bioch. Biophys. 135, 50.(With P.A. GURR, P.M. BRONSKILL & J.T-F. WONG) Purification of isoenzyme and coenzymes for kinetic study of horse liver alcohol dehydrogenase. Can. Biochem. 50, 1376.(With P.M. BRONSKILL, P.A. GURR & J.T-F. WONG). Kinetic mechanism for the major isoenzyme of horse liver alcohol dehydrogenase. Can. Biochem. 50, 1385.(With J.T-F. WONG, P.A. GURR & P.M. BRONSKILL) Heterotropic interactions within the reaction-centre of liver alcohol dehydrogenase. Reprinted from Analysis and Simulation of Biochemical Systems, FEBS, Amsterdam.(With P.A. GURR & J.T-F. WONG) Automated determination of enzymatic rate functions by gradient-flow method. Anal. Biochem. 51, 584.(With J.T-F. WONG) Topological analysis of enzymic mechanisms. (FEBS Advanced Course on Mathematical Models of Metabolic Regulation, November 8-16, Oberhof, German Democratic Republic.) Acta Biol. Med. 31, 507.(With P.A. COLLINS & J.T. WONG) Parallelism between ethanol and imidazole interactions with horse liver alcohol dehydrogenases, Canad. J. Biochem. 56, 1016-1020. In addition there were approximately 25 minor publications, especially reports of studies on fruits and vegetables, carried out as part of the program of the Food Investigation Board in the period 1934—1939.