Modification of a specific tyrosine enables tracing of the end‐to‐end distance during apomyoglobin folding

FEBS Letters

Poulsen, Flemming M.¹  Rischel, Christian¹ 

[1] Carlsberg Laboratorium, Kemisk Afdeling, Gamle Carlsberg Vej 10, DK-2500 Copenhagen Valby, Denmark

关键词: Protein folding;    Energy transfer;    Nitro-tyrosine;    Stopped-flow;    CD;    circular dichroism;    AEDANS;    5-(((acetylamino)ethyl)amino) naphthalene-1-sulfonic acid;    apoMb;    apomyoglobin;    Tyr(NO2);    3-nitro-tyrosine;    apoMb-Y146(NO2);    apoMb with Tyr146 converted to Tyr(NO2);    SDS-PAGE;    sodium dodecyl sulfate polyacryl-amide gel electrophoresis;    TNM;    tetranitromethane;    HPLC;    high performance liquid chromatography;    MALDI-TOF;    matrix assisted laser desorption ionization - time-of-flight (mass spectroscopy);    ACH;    α-cyano-4-hydroxy-cinnamic acid;    GuHCl;    guanidine-hydrochloride;   

DOI  :  10.1016/0014-5793(95)01087-U

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

In order to follow the overall geometry of the apomyoglobin molecule during folding, we have converted a specific tyrosine residue into 3-nitro-tyrosine. The specificity of the modification was verified by proteolytic cleavage of the modified protein and mass spectroscopy of the resulting fragments. By measuring the energy transfer from the tryptophanyl side-chains to the modified residue the average end-to-end distance can be followed. The experiment shows that after initiation of folding the N- and C-termini are rapidly brought into proximity, possibly to a near-native distance.

【 授权许可】

Unknown