Interdomain interactions within the gene 3 protein of filamentous phage

FEBS Letters

Riechmann, Lutz¹  Hartley, Oliver²  Winter, Greg²  Griffiths, Andrew D.²  Chatellier, Jean²  Fersht, Alan R.² 

[1] Laboratory of Molecular Biology, MRC Centre, Hills Road, Cambridge CB2 2QH, UK;Centre for Protein Engineering, MRC Centre, Hills Road, Cambridge CB2 2QH, UK

关键词: Phage fd;    Phage display;    Protein-ligand interaction;    Selection;    GroEL minichaperone;    g3p;    gene 3 protein;    D1;    N-terminal domain of g3p;    D2;    central domain of g3p;    D3;    C-terminal domain of g3p;    ELISA;    enzyme-linked immunosorbent assay;    GroEL*;    minichaperone GroEL(191–345);    SIP;    selective infection of phage;    PBS;    phosphate-buffered saline;    PEG;    polyethylene glycol;    cam;    chloramphenicol;    tet;    tetracycline;    kan;    kanamycin;    R;    resistance;    cfu;    colony-forming unit;   

DOI  :  10.1016/S0014-5793(99)01658-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Infection of Escherichia coli by filamentous phage fd is mediated by the phage gene 3 protein (g3p). The g3p consists of three domains (g3p-D1, D2 and D3) linked by flexible glycine-rich linkers. All three domains are indispensable for phage infectivity; the g3p-D1 domain binds to the TolA receptor presumably at the inner face of the outer membrane, the g3p-D2 domain to the F-pilus and the g3p-D3 domain anchors g3p to the phage coat. The N-terminal domains g3p-D1 and D2 interact with each other; this interaction is abrogated by binding of g3p-D2 to the F-pilus leading to the release of g3p-D1 to bind to TolA. Here, using phages with deletions in g3p, we have discovered a specific interaction between the two N-terminal domains and g3p-D3, the C-terminal domain of g3p. We propose that these interdomain interactions within g3p lead to a compact and stable organisation when displayed on the phage tip, but that during infection, this compact state must be unraveled.

【 授权许可】

Unknown