Screening of a library of phage-displayed peptides identifies human Bcl-2 as a taxol binding protein

被引:206
作者
Rodi, DJ
Janes, RW
Sanganee, HJ
Holton, RA
Wallace, BA
Makowski, L [1 ]
机构
[1] Florida State Univ, Inst Mol Biophys, Tallahassee, FL 32306 USA
[2] Univ London Queen Mary & Westfield Coll, Sch Biol Sci, London E1 4NS, England
[3] Florida State Univ, Dept Chem, Tallahassee, FL 32306 USA
[4] Univ London, Birkbeck Coll, Dept Crystallog, London WC1E 7HX, England
基金
美国国家科学基金会;
关键词
phage display; paclitaxel; Bcl-2; apoptosis;
D O I
10.1006/jmbi.1998.2303
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A random library of phage displayed peptides was screened for binding to a biotinylated derivative of paclitaxel (Taxol). Affinity-selected peptides were analyzed for similarity to human proteins. There was no significant similarity between the paclitaxel-selected peptides and tubulin. However, a subset of the peptides was identified that exhibits significant similarity to a non-conserved region of the anti-apoptotic human protein Bcl-2: ELISA assays confirmed binding of paclitaxel to Bcl-2, and circular dichroism spectroscopy demonstrated that a substantial conformational change accompanies this binding. In vivo, treatment with paclitaxel has been shown to lead to Bcl-2 inactivation with concomitant phosphorylation of residues in a disordered, regulatory loop region of the protein. Similarity between paclitaxel-selected peptides and this loop region implicate these residues in drug binding, and suggest that the apoptotic action of paclitaxel may involve the binding of paclitaxel to Bcl-2. These results demonstrate that peptides displayed on the surface of bacteriophage particles can mimic the ligand-binding properties of disordered regions of proteins. (C) 1999 Academic Press.
引用
收藏
页码:197 / 203
页数:7
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