Identifying and characterizing a second structural domain of protein disulfide isomerase

FEBS Letters

Kemmink, Johan¹  Penka, Elke¹  van Straaten, Monique¹  Darby, Nigel J.¹  Vincentelli, Renaud¹ 

[1] European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69012 Heidelberg, Germany

关键词: Disulfide bond;    Protein disulfide isomerase;    Protein domain;    Protein folding;    Thioredoxin;    CHAPS;    3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate;    DTT;    1;    4-dithiothreitol;    HPLC;    high pressure liquid chromatography;    IPTG;    isopropyl-β-d-thiogalactopyranoside;    PAGE;    polyacrylamide gel electrophoresis;    PDI;    protein disulfide isomerase;    PDO;    protein disulfide oxidoreductase;    PMSF;    phenyl-methylsulfonylfluoride;    TLCK;    N α-p-tosyl-l-lysine chloromethylketone;    TFA;    trifluoroacetic acid;    PDI i–j;    polypeptide construct corresponding to residues i to j using the residue numbering of mature human PDI;   

DOI  :  10.1016/S0014-5793(99)00374-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Recent protein engineering studies have confirmed the multidomain nature of protein disulfide isomerase previously suggested on the basis of analysis of its amino acid sequence. The boundaries of three domains, denoted a, a′ and b, have been determined, and each domain has been expressed as an individual soluble folded protein. In this report, the boundaries of the final structural domain, b′, are defined by a combination of restricted proteolysis and protein engineering approaches to complete our understanding of the domain organization of PDI. Using these data an optimized polypeptide construct has been prepared and characterized with a view to further structural and functional studies.

【 授权许可】

Unknown