Selection of peptides that bind to plasminogen activator inhibitor 1 (PAI‐1) using random peptide phage‐display libraries

FEBS Letters

Pannekoek, Hans¹  Holm, Arne³  Gårdsvoll, Henrik¹  van Zonneveld, Anton-Jan¹  Danø, Keld²  van Meijer, Marja¹  Eldering, Eric¹ 

[1] Department of Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 15, 1105 AZ Amsterdam, The Netherlands;The Finsen Laboratory, Rigshospitalet, Strandboulevarden 49, DK-2100 Copenhagen, Denmark;Chemical Department, Royal Veterinary and Agricultural University, Thorvaldvej 40, DK-1871 Copenhagen, Denmark

关键词: Phage display;    Plasminogen activator inhibitor 1;    Peptide;    Peptide library;    BSA;    bovine serum albumin;    ELISA;    enzyme-linked immunosorbent assay;    LB;    Luria broth;    PAI-1;    plasminogen activator inhibitor 1;    SMB;    somatomedin B;    TBS;    Tris-buffered saline;    t-PA;    tissue-type plasminogen activator;    u-PA;    urokinase-type plasminogen activator;   

DOI  :  10.1016/S0014-5793(98)00742-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Large random hexa- and decapenta-peptide libraries were constructed and displayed on the surface of the filamentous phagemid pComb8. Panning of the hexa-peptide library on immobilized plasminogen activator inhibitor 1 (PAI-1) specifically selected a minor fraction of concatemers, indicating that binding to PAI-1 requires an extended amino acid sequence. Accordingly, the decapenta-peptide library exclusively yielded PAI-1 binding peptides of 15 amino acid residues. None of these phage-bound peptides prevented the interaction between PAI-1 and its target serine protease urokinase (u-PA). To isolate peptides that block the interaction between PAI-1 and u-PA, phages bound to immobilized PAI-1 were eluted by incubation with u-PA. Remarkably, this procedure resulted in elution of a unique phage type that harbors a concatemer of decapentamers, consisting of 49 amino acid residues with no obvious similarity to the primary sequence of PAI-1 or u-PA.

【 授权许可】

Unknown