Protein sequencing

Details Protein sequencing Protein sequencing

1991-06-21

1993-12-14

Housel, James C.

Chemistry: analytical and immunological testing

Peptide, protein or amino acid

Amino acid or sequencing procedure

436 57, 436172, 530337, 530343, 530407, 530802, G01N 2164, G01N 3368

Patent

active

052702138

ABSTRACT:
A method for the generation of phenylthiocarbamyl (PTC) amino acids from phenylthiohydantoin (PTH) or anilinothiozolinone (ATZ) amino acids. The method involves a base-catalyzed ring opening of the PTH or ATZ in the presence of a reducing agent. The method affords an alternative to the established aqueous acid conversion reaction of the Edman degradation in which ATZ and PTC amino acids are converted to the PTH amino acid. A further method is described for the generation of reactive ATZ amino acids from PTH amino acids. These methods facilitate the analysis of protein at low molar amounts by allowing the synthesis of amino acid derivatives which can be analyzed in quantities which are much lower than those required for conventional PTH amino acid analysis.

REFERENCES:
patent: 5051369 (1991-09-01), Tsugita et al.
A. Tsugita, et al; "Sensitization of Edman Amino Acid Derivatives Using the Fluroescent Reagent, 4-Aminofluorescein"; The Journal of Biochemistry; vol. 106, No. 1, Jul. 1989, pp. 60-65.
Chemical Abstract; vol. 111, No. 11, 11 Sep. 1989, Columbus, Ohio, US; abstract No. 93162.
Chemical Abstract; vol. 84, No. 17, 28 Apr. 1986, Columbus, Ohio, US, abstract No. 147104e.
Edman, P. and A. Begg, "A Protein Sequenator," European Journal of Biochemistry, 1967, vol. 1, p. 80.
Matsudiara, P., "Sequence from Picomole Quantities of Proteins Blotted onto Polyvinyliene-difluoride Membranes," Journal of Biological Chemistry, 1987, vol. 262, pp. 10035-10038.
Inman, J. K. and E. Apella, "Identification of Anilinothiozolinones after Rapid Conversion to N-Phenylthiocarbamyl-Amino Acid Methylamides," Methods in Enzymology, Academic Press, Inc., 1977, vol. 47, pp. 374-385.
Tsugita et al., "Sensitization of Edman Amino Acid Derivatives Using the Fluorescent Reagent, 4-Aminofluorescein," Journal of Biochemistry, 1989, vol. 106, pp. 60-65.
Horn et al., "Sensitization of Edman Amino Acid Derivatives for Sensitivity Enhancement in Edman Degradation," Techniques in Protein Chemistry, Academic Press, Inc., 1989, pp. 51-59.
Tarr, G., "Manual Edman Sequencing System," Methods of Protein Microcharacterization, The Humana Press, Inc., 1986, pp. 155-194.

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