A new strategy for primary structure determination of proteins: Application to bovine β‐casein

FEBS Letters

Ribadeau-Dumas, Bruno²  Carles, Christophe²  Huet, Jean-Claude¹ 

[1] Institut National de la Recherche Agronomique, CNRA, 78000 Versailles, France;Institut National de la Recherche Agronomique, CRJ, 78350 Jouy-en-Josas, CNRA, 78000 Versailles, France

关键词: Protein;    Sequence determination;    β-Casein;    (Bovine);    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    TFA;    trifluoroacetic acid;    IBA;    iodosobenzoic acid;    TEA;    triethylamine;    PITC;    phenylisothiocyanate;    FPLC;    fast protein liquid chromatography;    RP-HPLC;    reverse-phase high-performance liquid chromatography;    PMSF;    phenylmethanylsulfonyl fluoride;    CPD-A;    carboxypeptidase A;    CPD-Y;    carboxypeptidase Y;    ATZ;    anilinothiazolinone;   

DOI  :  10.1016/0014-5793(88)81138-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A new approach has been developed for sequencing proteins. A radioactive label is attached specifically to the C-terminus of the protein. The labelled molecule is subjected to varying proteolysis conditions. From the electrophoretic patterns (SDS-PAGE) of the hydrolysates, appropriate cleavage conditions are selected, giving labelled peptides of different lengths which are purified. The labelled peptides are sequenced in order of increasing size (from 1 to n), peptide (i) being sequenced until the N-terminal sequence of peptide (i-1) is encountered. This approach allows the determination of a complete protein sequence with a minimal number of Edman cycles. The method was successfully applied to bovine β-casein (209 residues) which was completely resequenced with only 239 Edman cycles.

【 授权许可】

Unknown