Sequence determination, proteins

The unstable thiazolinones are converted into stable hydantoines in order to facilitate their identification. Conversion and identification are carried out outside the instrument after extraction of the thiazolinones with butylchlor-ide. The conversion reaction as well as the problems associated with identification of the PTH-amino acids were studied in detail by Edman and described explicitly in Needleman s book on Protein Sequence Determination Conversion is generally carried out in 1 N HCl at 80 °C within 10 min. The PTH-derivatives are extracted from the aqueous phase with ethyl acetate with the exception of PTH-arginine, PTH-histidine and PTH-cysteine which remain in the aqueous phase. 

Edman, P., in Protein Sequence Determination (ed. S.B. Needleman), p. 211. Ber-lin-Heidelberg-New York Springer 1970. 

The covalent structure of insulin was established by Frederick Sanger in 1953 after a 10-year effort. This was the first protein sequence determination.237 238 Sanger used partial hydrolysis of peptide chains whose amino groups had been labeled by reaction with 2,4-dinitrofluorobenzene239 to form shorter end-labeled fragments. These were analyzed for their amino acid composition and labeled and hydrolyzed again as necessary. Many peptides had to be analyzed to deduce the sequence of the 21-residue and 30-residue chains that are joined by disulfide linkages in insulin.237 238... 

GLC is an important adjunct to protein sequence determination. Automatic "sequenators" based upon the approach developed by Edman are available and have been described in detail by Niall (60). The Edman degradation, summarized in Equation 9.5, makes use of methyl or phenylisothiocyanate which reacts with the N-terminus of a peptide. Exposure of the isothiocyanate derivative of the protein to acid results in cleavage of the terminal amino acid as a thiaxolinones and exposure of the next amine group on the peptide. Thus, the process can be repetitively carried out, each amino acid removed from the peptide, in a sequential manner. Thiazolinones rearrange in acid medium to form thiohydantoin derivatives of amino acids, some of which may be directly gas chromatographed others must be derivatized typically as trimethylsilyl derivatives. 

Dayhoff, M. 0. Computer aids to protein sequence determination. J. Theoret. Biol. 8, 97 (1964). 

Protein sequence determination uses the standard sequential Edman-degradation procedure to cleave of the N-terminal amino acid of the protein followed by the characterization of the released amino acid in each... 

Protein sequencing . Determine the sequence of hexapeptide based on the following data. Note When the sequence is not known, a comma separates the amino acids. (See Table 4.3)... 

Edman, P, (1970) Sequence determination. In Needleman, S. B., ed.. Protein Sequence Determination (Springer-Verlag, Berlin) p. 211,... 

Pisano, J. J., Analysis of amino acid phenylthiohydantoins by gas chromatography and high-performance liquid chromatography, in Protein Sequence Determination, Needleman, S. B., Ed. Springer-Verlag, New York, also listed as Mol. Biol. Biochem. Biophys., 8, 280-296, 1975. 

Chauvet, S., Merabet, S., Bilder, D., Scott, M.P., Pradel, J., Graba, Y. 2000. Distinct hox protein sequences determine specificity in different tissues. Proc. Natl. Acad. Sci. USA 97, 4064-4069. 

P, Edman in S.B. Needleman, Protein Sequence Determination, S. 211, Chapman z Hall, London... 

Gish, D. in Protein Sequence Determination. A Sourcebook of Methods... 

S. Blackburn, Protein Sequence Determination, Marcel Dekker, New York, 1970. 

T. L. Blundell and L. N. Johnson, Protein Crystallography, Academic Press, New York, 1976. S. B. Needleman (ed.). Protein Sequence Determination, second ed.. Springer, New York, 1975. S. Blackburn (ed.). Amino Acid Analysis, second ed., Marcel Dekker, New York, 1978. 

Vol. 8 Protein Sequence Determination. A Sourcebook of Methods and Techniques. Edited by Saul B. Needleman. With 77 figures. XXI, 345 pages. 1970. DM 84,— US 34.50... 

ScoFFONE, E., and A. Fontana Identification of Specific Amino Acid Residues. Protein Sequence Determination. Methods and Techniques (S. B. Needleman, Ed.), p. 185-210. New York Springer. 1970. 

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