Automated sequence analyzer

Edman, Pehr Victor, 1916-1977 (pp. 118,240, Plate 15) born in Stockholm in 1916, matriculation examination in 1935, studied medicine at the Karolinska Institute Medical School in Stockholm from 1935, Bachelor of Medicine in 1938, graduation as a physician in 1946. Concurrently with his studies in medicine he started his training in biochemistry with Erik Jorpes, for a short time also with Hugo Theorell, and soon started a project on angiotensin that led to a MD thesis. Then he widened his experience in protein chemistry during one year at the Rockefeller Institute in Princeton with Northrop and Kunitz (crystallization of proteolytic enzymes). On his return to Sweden, Edman was awarded an associate professorship in Lund in 1947 where he conducted his stepwise peptide degradation work (p. 118) between 1950 and 1956. In 1957 Pehr Edman accepted an offer to be Director of Research at St. Vincent s School of Medical Research in Melbourne, Australia, where he remained for 15 years, during which the work on an automated sequence analyzer was finished in 1967. From 1972 until his death from a brain tumor in 1977 he was Director of the Department of Protein Chemistry of the Max-Planck-Institute for Biochemistry in Martinsried near Munich. 

A blank control run was performed by injection of the above buffer. Then, at the same eluent composition, vacancy peaks were obtained by injection of the above sample solutions containing only HSA in the buffer. Blank and protein injections were run using the eluents containing different drug concentrations (0.4-50 pM) in an automated sequence, controlled by the workstation. The experimental data obtained by the modified Hummel-Dreyer method were analyzed using Scatchard plots. Binding affinity was calculated from a Langmuir-type equation. 

To determine the structure of a protein or peptide, we need to answer three questions What amino acids are present How much of each is present in what sequence do the amino acids occur in the peptide chain The answers to the first two questions are provided by an automated instrument called an amino acid analyzer. 

Protein Sequencing. The xylanase protein sequence was obtained by stepwise automated E an degradation ter an initial double-coupling protocol, using an Applied Biosystems model 470A gas-phase sequenator equipped with an on-line Applied Biosystems model 120A PTH-amino acid analyzer 13). 

These techniques have been used successfully in the micro-Zdman degradation of the enzyme mouse sarcoma dihydrofolate reductase to obtain the amino acid sequence of the first 25 amino acids 455). Similarly, RPC has been used in coqjunction with the automated Edman technique for sequencing 32 residues of myoglobin 456). Methionine and its oxidation products, methionine sulfoxide and methionine sulfone, in methionine fortified foods have been analyzed as their dansyl derivatives 457). Lysine has been determined as its dansyl derivative in a study in which the stability of lysine in fortified wheat flour was evaluated (458). 

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