Solid‐phase Edman degradation. The use of p‐phenyl diisothiocyanate to attach lysine‐ and arginine‐containing peptides to insoluble resins

作者：Richard A. Laursen, Marcus J. Horn, Alex G. Bonner · 发表于：FEBS Letters · 年份：1972 · DOI：10.1016/0014-5793(72)80165-0 · 被引用次数：136 · 研究领域：RNA and protein synthesis mechanisms、Chemical Synthesis and Analysis、RNA Interference and Gene Delivery

The primary step in sequencing peptides by the solid-phase Edman degradation [l] is attachment of the peptide to an insoluble resin, generally by blocking the peptide amino groups and then activating the C-terminal carboxyl with carbonyldiimidazole [l] or a carbodiimide [2-41. A problem limiting the usefulness of the solid-phase method is that the side-chain carboxyls of aspartic and glutamic acids also become activated, with the result, at least when carbonyldiimidazole is used, that the y-carboxyl of glutamic acid becomes bound to the resin and that aspartic acid apparently forms a cyclic imide which prevents further degradation of the peptide [i] . Although procedures for selective blocking of side chain carboxyls have been devised [2], they involve a number of steps and are often unsuitable for use with nanomole amounts of peptide. A procedure we have found to be particularly useful for tryptic peptides involves attaching them by their lysine e-amino groups to aminopolystyrene usingp-phenyl diisothiocyanate. Arginine peptides are first deguanidated with hydrazine [5],