タンパク質分析 : 試薬の開発と応用

概要

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Reagents and techniques for the analysis of primary structures of proteins have been reviewed with special emphasis on those which had been developed by the present author and his coworkers : Fluorescent Koshland reagent (FKR, 2-carboxy-1-hydroxy-4-naphtylmethyldimethylsulfonium chloride) for tryptophan residues and fluorescent Edman reagent (4-N, N-dimethylainino-l-naphthyl isothiocyanate, DNTC). FKR reacts with tryptophan residues at pH 03 and room temperature to produce fluorescent adduct (λex255 nm, λem 416 nm), the reaction being completed within 2 min. Owing to its high specificity, FKR was applicable to the determination of tryptophan residues in peptides and proteins. Methods for the selective isolation of tryptophan-containing fragments in peptides and proteins, based upon chelating ability of FKR derivative and reversed-phase high performance liquid chromatography, are described. Scope and limitation of DNTC in microsequence analysis of proteins are discussed, together with other Edman-type reagents. Furthermore, the reagents for protein assay and amino acid analysis are briefly reviewed.
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