Robert Bruce Merrifield ( born Robert Bruce Merrifield ; July 15, 1921 , Fort Worth , Texas - May 14, 2006 , Crescill , NJ ) is an American biochemist, 1984 Nobel Prize in Chemistry for his proposed method of chemical synthesis on solid matrices [1] .
| Robert Bruce Merrifield | |
|---|---|
| English Robert Bruce Merrifield | |
 | |
| Date of Birth | July 15, 1921 |
| Place of Birth | Fort Worth , Texas , USA |
| Date of death | May 14, 2006 (84 years old) |
| Place of death | Crescill , New Jersey , USA |
| A country |  USA |
| Scientific field | biochemistry |
| Place of work | |
| Alma mater | |
| Awards and prizes |  Nobel Prize in Chemistry (1984) |
Content
Childhood and Youth
Bruce was born July 15, 1921 in Fort Worth , Texas. He was the only child in the family. His father, George E. Merrifield, was an interior designer and sold hardware, his mother was Lorene née Lucas.
In 1923, when Bruce was two years old, his family moved to Southern California. There was a time of the Great Depression , and the Merrifilds, in order to find a job for their father, had to move from place to place. Because of this, Bruce often changed schools.
His favorite subjects were physics , chemistry and astronomy . Bruce especially liked to conduct chemical experiments. After graduating from a California primary school (grade 9), Merrifield went to high school at the Montebello High School in 1937-1939. After two years at Pasadena Junior College, he transferred to the University of California at Los Angeles (UCLA). Having received a bachelor's degree there in 1943, Bruce worked for about a year at the Philip R. Park Research Foundation, assisting in experiments with feeding animals with synthetic amino acids. One of these experiments was the Geiger experiment, which first showed the need for the joint presence of essential amino acids in food for the normal development of animals.
Career start
A year later, Bruce entered graduate school at the University of California and in 1944 began dissertation research under the direction of professor of biochemistry Max Dunn (MS Dunn). The latter studied amino acids and suggested Merrifield to tackle this topic. Bruce began to work with biopolymers that affect the growth of bacteria of the genus lactobacilli , which were used for the quantitative analysis of amino acids. In this area, Dunne and Merrifield were pioneers. Bruce determined proline and also studied pyrimidine bases containing in yeast. For the development (together with his supervisor Dunn) of a biochemical method for the quantitative analysis of pyrimidines, he received his doctorate (Ph.D.) in 1949.
Immediately after graduation, on June 19, 1949, he married Elizabeth Furlong and left for New York the next day.
There, under the leadership of Dr. Woolley (DW Woolley), Bruce began working as an assistant biochemist at the Rockefeller Institute for Medical Research (later Rockefeller University ). He investigated a new bacterial growth factor, strepogenin, which his supervisor discovered. By this time, Sanger had determined the primary structure of insulin, and Merrifield, thanks to the high activity of strepogenin, was able to study two peptides from the insulin B chain, the pentapeptide and heptapeptide containing serine [2] .
Solid Phase Peptide Synthesis Method
By the middle of the 20th century, the most common method for the synthesis of peptides was the method of Emil Fischer , who was the first to begin work in this area. In this method, amino acids are added sequentially to the resulting peptide chain. Cycles consisting of the stages of group protection, activation, attachment to the peptide chain and removal of the protective group are repeated until a peptide chain of the desired composition is obtained. In the synthesis process, a large number of by-products formed, which had to be removed at each stage. As a rule, the yield of the target peptide synthesized by this method was very small, and the product itself was heavily contaminated.
Some improvements in this process were developed in the 1930s. One such improvement was the use of the universal benzyloxycarbonyl group, which allowed the Vincent du Vigno group to synthesize oxytocin and vasopressin .
After synthesizing peptides for several years by methods known at that time, Bruce began to think about creating an alternative, wider version of the syntheses. In his laboratory journal in May 1959, he wrote:
| There is a need for a fast, quantitative and automatic method for the synthesis of long chain peptides. A possible approach may be the use of chromatographic columns in which activated amino acids are added to the peptide attached to the polymer carrier, then the protective groups are removed and then the cycle is repeated until the desired peptide is built. In the last step, the peptide must be removed from the carrier |
After a series of experiments, Bruce decided to use styrene and divinylbenzene polymer as a carrier. This version of the carrier was attractive as its high solubility in organic solvents, and a low degree of crosslinking. To the advantages of this carrier, one can add that it is easy to modify it so that the first amino acid is attached to the polymer in the form of a benzyl ester.
Among the few protective groups available by 1960, the benzyloxycarbonyl group for the new variant of peptide synthesis was not entirely suitable, since it required a strongly acidic environment to separate from the product, and then Bruce decided to use dicyclohexylcarbodiimide (DCCI) in his experiments, which was used shortly before .
In 1963, Merrifield published a classic article in the Journal of the American Chemical Society , in which he describes a method he called “solid phase peptide synthesis” [3] . This article has become one of the most cited publications in the journal.
An important step forward was the design of a plant capable of automating the synthesis of peptides. The first working device was made by Bruce and his assistant John Stuart with the assistance of N. Jernberg in 1965.
The publication of Merrifield's work describing the preparation of a tetrapeptide in a new setup presented solid-phase peptide synthesis (SPPS) as an alternative to peptide synthesis [4] . Although skeptical scientists were found, the article attracted enormous attention of chemists around the world. Soon after the article on the tetrapeptide, a series of publications followed devoted to a detailed description of the new method [5] . The group in which Bruce worked, using the equipment they developed, also received several peptide hormones, including bradykinin , oxytocin , angiotensin, and insulin protein [6] . With the spread of the solid-phase peptide synthesis method, commercial models of automated synthesizers began to appear on the market, many of which were of poor quality and strongly discredited SPPS.
Opponents of the new method for the synthesis of peptides argued that the peptide products obtained using an automated synthesizer were not pure. Merrifield faced the challenge of product purity throughout its peptide synthesis work. Its solution was made possible thanks to the development of high performance liquid chromatography .
Ribonuclease Synthesis
In 1969, Merrifield, together with Bernd Gutte, successfully synthesized the ribonuclease enzyme [7] . To carry out this synthesis, Merrifield and Gutta needed 369 chemical reactions and 11,931 separate stages, which required several weeks of continuous operation of an automated synthesizer. This work served as an experimental proof of the Anfinsen hypothesis, which suggested that the primary structure of a protein determines its tertiary structure.
Synthesis of cecropin and its analogues
In 1982, Bruce Merrifield and colleagues first synthesized the peptide cecropin , which has an antibacterial effect. Studies of this substance subsequently turned into a separate area with a systematic classification of this class of compounds [8] . Some representatives from this class were successfully synthesized and analyzed for antibacterial activity in the laboratory where Bruce worked [9] . During the experiments, scientists found that the D-cecropins A obtained by them are able to create the same electrical conductivity as their enantiomers in the bilipid layer, and also have similar antimicrobial activity [10] [11] [12]
In addition, it was found that cecropins in non-polar media have a high degree of spiralization. Based on the results, Bruce Merrifield and his laboratory staff concluded that these peptides act through the formation of ion channels in cell membranes and do not interact with chiral centers like enzymes and receptors.
The Merrifield method stimulated great progress in biochemistry , pharmacology and medicine , making it possible to systematically study the dependence of the activity of enzymes, hormones and antibodies on their structure. The development and improvement of the method was the main research topic in his laboratory, which he directed until the last days of his life.
Prizes and Honors
For his work, Merrifield was elected a member of the US National Academy of Sciences in 1972 [13] .
He was awarded a number of prizes for his work on the synthetic chemistry of peptides, including:
- Lasker Prize (1969)
- Geirdner International Award (1970)
- American Chemical Society Award for Key Work in Synthetic Organic Chemistry (1972)
- Nichols Medal (1973)
- American Peptide Society Alan Pierce Award (1979)
- Nobel Prize in Chemistry (1984)
- Century Award (1986)
- Ralph F. Hirschmann Prize of the German Peptide Society (1990).
Personal life
Bruce was married to Elizabeth Farlong (1949), whom he met at the University of Los Angeles. After receiving a biological education, she worked with her husband at Rockefeller University for over 20 years. In 1993, he published his autobiography, Life in the Golden Age of Peptide Chemistry. After a long illness, R. Bruce Merrifield died on May 14, 2006, at the age of 84, at his home in Cresskill, New Jersey. He left behind his wife, 6 children (Nancy, Jim, Betsy, Katie, Lori, and Sally) and 16 grandchildren.
Literature
- Ragnarsson U. Bruce Merrifield (1921-2006) // National Academy of Sciences. - 2013.
- Jones, John H (2007), “ RB Merrifield: European footnotes to his life and work ”, Journal of Peptide Science T. 13 (6): 363–7, 2007 Jun, PMID : 17516587, doi : 10.1002 / psc.857 , < https://www.ncbi.nlm.nih.gov/pubmed/17516587 >
- Gierasch, Lila M (2006), " Editorial: Passing of a gentle giant of peptide science: in memoriam, R. Bruce Merrifield ", Biopolymers T. 84 (5): 433-4, PMID : 16850497, doi : 10.1002 / bip .20574 , < https://www.ncbi.nlm.nih.gov/pubmed/16850497 >
- Lerner, Richard A (2006), Retrospective. R. Bruce Merrifield (1921-2006) , Science T. 313 (5783): 57, 2006 Jul 7, PMID : 16825560, PMID 16825560 , doi : 10.1126 / science.1130957 , < https://www.ncbi.nlm.nih.gov/pubmed/16825560 >
- Sano, S (1985), " [Work of Dr. Robert Merrifield, with special reference to the development of solid phase peptide synthesis ]", Tanpakushitsu Kakusan Koso T. 30 (1): 51–3, Jan Jan, PMID : 3883428 , < https://www.ncbi.nlm.nih.gov/pubmed/3883428 >
- Domagk, GF (1984), " [The 1984 Nobel prize for chemistry. Synthetic hormones and enzymes with Merrifield's peptide synthesizer ]", Dtsch. Med. Wochenschr. T. 109 (49): 1901–2, 1984 Dec 7, PMID : 6389076 , < https://www.ncbi.nlm.nih.gov/pubmed/6389076 >
Notes
- ↑ Kenneth Chang . R. Bruce Merrifield, Who Won Nobel Prize in Chemistry, Dies at 84 , The New York Times (May 6, 2006). Date of appeal April 12, 2008. (English)
- ↑ RB Merrifield, DW Woolley. Isolation and determination of structure of a peptide with strepogenin activity (English) // Archives of Biochemistry and Biophysics : journal. - Elsevier , 1955. - Vol. 56 , no. 1 . - P. 265-266 . - DOI : 10.1016 / 0003-9861 (67) 90184-1 .
- ↑ RB Merrifield. Solid Phase Peptide Synthesis. I. The Synthesis of a Tetrapeptide (Eng.) // Journal of the American Chemical Society : journal. - 1963. - Vol. 85 . - P. 2149 . - DOI : 10.1021 / ja00897a025 .
- ↑ RB Merrifield. Automated Synthesis of Peptides (Eng.) // Science. - 1965. - P. 178-185 . - DOI : 10.1126 / science.150.3693.178 .
- ↑ Robert B. Merrifield,, John Morrow. Stewart,, Nils. Jernberg. Instrument for automated synthesis of peptides (neopr.) // Analytical Chemistry. - 1966. - S. 1905-1914 . - DOI : 10.1021 / ac50155a057 .
- ↑ B. Marglin, RB Merrifield. The Synthesis of Bovine Insulin by the Solid Phase Method (English) // Journal of the American Chemical Society : journal. - 1966. - P. 5051-5052 . - DOI : 10.1021 / ja00973a068 .
- ↑ RB Merrifield, B. Gutte. The Synthesis of Ribonuclease A (Eng.) // The Journal of Biological Chemistry : journal. - 1971. - Vol. 246 . - P. 1922-1941 .
- ↑ Håkan Steiner, David Andreu, RB Merrifield. Binding and action of cecropin and cecropin analog: Antibacterial peptides from insects // Biochimica et Biophysica Acta : journal. - 1988. - Vol. 939 , no. 2 . - P. 260-266 . - DOI : 10.1016 / 0005-2736 (88) 90069-7 .
- ↑ HG Boman, I. Faye, P. Hofsten, K. Kockum, J.-Y. Lee, KG Xanthopoulos, H. Bennich, Å. Engström, RB Merrifield, D. Andreu. On the primary structures of lysozyme, cecropins and attacins from Hyalophora cecropia (Eng.) // Developmental & Comparative Immunology: journal. - 1985. - Vol. 9 , no. 3 . - P. 551–558 . - DOI : 10.1016 / 0005-2736 (88) 90069-7 .
- ↑ HG Boman, D. Wade, IA Boman, B. Wåhlin, RB Merrifield. Antibacterial and antimalarial properties of peptides that are cecropin-melittin hybrids (English) // FEBS Letters : journal. - 1989. - Vol. 259 , no. 1 . - P. 103-106 . - DOI : 10.1016 / 0014-5793 (89) 81505-4 .
- ↑ David Andreu, Josep Ubach, Anita Boman, Birgitta Wåhlin, David Wade, RB Merrifield, Hans G. Boman. Shortened cecropin A-melittin hybrids Significant size reduction retains potent antibiotic activity (Eng.) // FEBS Letters : journal. - 1992. - Vol. 296 , no. 2 . - P. 190—194 . - DOI : 10.1016 / 0014-5793 (92) 80377-S .
- ↑ Francesca M. Marassi, Stanley J. Opella, Padmaja Juvvadi, RB Merrifield. Orientation of Cecropin A Helices in Phospholipid Bilayers Determined by Solid-State NMR Spectroscopy (Eng.) // Biophysical Journal : journal. - 1999. - Vol. 77 , no. 6 . - P. 3152-3155 . - DOI : 10.1016 / S0006-3495 (99) 77145-6 .
- ↑ Bruce Merrifield