Multidrug-resistance drug-binding peptides generated by using a phage display library.
A phage display library of random decapeptides was used to generate peptide ligands that can bind multidrug-resistance (MDR) drugs mimicking, in this respect, the drug-binding activity of P-glycoprotein. Seven peptide sequences were identified that specifically bound doxorubicin. Five of these sequences expressed the core consensus motif WXXW. The displacement assay showed that the phages expressing these peptides bound MDR type drugs (vinblastine, doxorubicin, verapamil, and genistein) with the same selectivity as P-glycoprotein and did not interact with non-MDR type drugs, such as arabinosylcytosine (Ara-C) and melphalan. One of the selected peptides that showed a highest capacity for the binding (VCDWWGWGIC) was synthesized and displayed competition with the phage for doxorubicin binding. The structure modeling suggested that all the selected sequences contained a hydrophobic envelope in which MDR drugs could be docked with substantial energy minimization. Western blot analysis showed that monospecific antibody obtained against the phage expressing VCDWWGWGIC peptide could specifically recognize P-glycoprotein in the membrane fraction of MDR phenotype MCF-7ADR cells. The MDR drug-binding sequences generated during this work could provide an important tool for design and screening of new chemotherapeutic agents.
Popkov
M
M
Institute Armand-Frappier, University of Quebec, Laval, Canada.
Lussier
I
I
Medvedkine
V
V
Estève
P O
PO
Alakhov
V
V
Mandeville
R
R
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000276:immunology* / Q000378:metabolism; D001435:Bacteriophages / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001943:Breast Neoplasms / Q000276:immunology / Q000378:metabolism; D004317:Doxorubicin / Q000378:metabolism*; D018432:Drug Resistance, Multiple; D015723:Gene Library; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D011487:Protein Conformation; D014407:Tumor Cells, Cultured
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D010455:Peptides; D004317:Doxorubicin