DOI: 10.1111/j.1432-1033.1971.tb01366.x. Solid‐Phase Edman Degradation

10.1111/j.1432-1033.1971.tb01366.x

Crossref journal-article

Wiley

European Journal of Biochemistry (311)

Abstract

A modification of the Edman degradation is described whereby peptides are degraded sequentially on a solid support by a machine, the solid‐phase peptide sequencer. Reagents are pumped through a column containing the resin‐bound peptide, and the amino acid thiazolinones released after each degradative cycle are collected on a fraction collector for subsequent analysis. A complete cycle requires 2 h, and all operations are controlled automatically by a programmer. An improved peptide resin is described, to which peptides can be attached in yields ranging from about 70% for the 30 amino acid insulin B‐chain to nearly 100% for smaller peptides. Of the 18 amino acids examined so far, glycine, alanine, phenylalanine, tyrosine, valine, leucine, isoleucine, proline, methionine, asparagine, glutamine, histidine, lysine, threonine, serine and cysteic acid are degraded normally; the phenylthiohydantion of threonine is partially dehydrated and that of serine is almost completely destroyed by trifluoroacetic acid, however. The phenylthiohydantion of glutamic acid is not detected, because it remains bound by its γ‐carboxyl group to the resin. When aspartic acid is encountered in peptides, further degradation is completely blocked, presumably as a result of the β‐carboxyl group having formed a cyclic imide. Using the sequencer and unpurified reagents, it was possible to degrade entirely a 0.24 μmole sample of the 21 amino acid insulin A‐chain (oxidized) and to analyze the products within a 3‐day period. Degradative yields averaged 94% per cycle. Similarly, 0.1 μmole of the insulin B‐chain was degraded through 18 cycles before identification of the phenylthiohydantoins became ambiguous. The sequencer is simple in design and operation, and is inexpensive to construct. The scope and limitations of the method are discussed.

Bibliography

Laursen, R. A. (1971). SolidâPhase Edman Degradation. European Journal of Biochemistry, 20(1), 89â102. Portico.

Authors 1

Richard A. Laursen (first)

References 26 Referenced 350

10.3891/acta.chem.scand.04-0277
10.1111/j.1432-1033.1967.tb00047.x
10.1021/ja00974a069
10.1002/zfch.19670070517 / Z. Chem. by Schellenberger A. (1967)
10.1002/recl.19670860110
10.1021/bi00897a032
{'key': 'e_1_2_1_8_2', 'first-page': '727', 'volume': '29', 'author': 'Laursen R. A.', 'year': '1970', 'journal-title': 'Fed. Proc.'} / Fed. Proc. by Laursen R. A. (1970)
10.1021/ja01502a038
10.1016/0006-291X(69)90862-6
{'key': 'e_1_2_1_11_2', 'first-page': '430', 'volume-title': 'Thin‐Layer Chromatography', 'author': 'Stahl E.', 'year': '1965'} / Thin‐Layer Chromatography by Stahl E. (1965)
10.1021/bi00863a030
10.1021/bi00840a011
Mross G. andDoolittle R. personal communication 1970.
10.1071/CH9630411
10.1021/ac50155a057
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10.1016/S0021-9258(18)63603-8
{'key': 'e_1_2_1_19_2', 'first-page': '728', 'volume': '29', 'author': 'Barnes W. T.', 'year': '1970', 'journal-title': 'Fed. Proc.'} / Fed. Proc. by Barnes W. T. (1970)
10.1016/0003-2697(68)90122-X
10.1021/bi00806a016
10.1042/bj1070728
10.1016/S0076-6879(67)11053-7
Horn M. unpublished observations 1970.
{'key': 'e_1_2_1_25_2', 'first-page': '728', 'volume': '29', 'author': 'Hermodson M. A.', 'year': '1970', 'journal-title': 'Fed. Proc.'} / Fed. Proc. by Hermodson M. A. (1970)
{'key': 'e_1_2_1_26_2', 'volume-title': 'Atlas of Protein Sequence and Structure', 'author': 'Dayhoff M. O.', 'year': '1969'} / Atlas of Protein Sequence and Structure by Dayhoff M. O. (1969)
10.1038/2201300a0

Dates

Type	When
Created	20 years, 5 months ago (March 3, 2005, 9:05 a.m.)
Deposited	1 year, 9 months ago (Nov. 23, 2023, 7:49 a.m.)
Indexed	11 months ago (Sept. 19, 2024, 11:01 a.m.)
Issued	54 years, 3 months ago (May 1, 1971)
Published	54 years, 3 months ago (May 1, 1971)
Published Online	20 years, 5 months ago (March 3, 2005)
Published Print	54 years, 3 months ago (May 1, 1971)

Funders 0

None

BibTeX

@article{Laursen_1971, title={Solid‐Phase Edman Degradation: An Automatic Peptide Sequencer}, volume={20}, ISSN={1432-1033}, url={http://dx.doi.org/10.1111/j.1432-1033.1971.tb01366.x}, DOI={10.1111/j.1432-1033.1971.tb01366.x}, number={1}, journal={European Journal of Biochemistry}, publisher={Wiley}, author={Laursen, Richard A.}, year={1971}, month=may, pages={89–102} }

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