Identification by Phage Display of the Linear Continuous MRPr1 Epitope in the Multidrug Resistance-Associated Protein (MRP1)
| Autor: | Stefano Barca, Franco Felici, Vito Mennella, Michela Flego, Maria Luisa Dupuis, Alessandro Ascione, Franca Moretti, Francesca Poloni, Maurizio Cianfriglia |
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| Rok vydání: | 2003 |
| Předmět: |
Phage display
Protein family medicine.drug_class Molecular Sequence Data Clinical Biochemistry DNA Single-Stranded Biology Monoclonal antibody Peptide Mapping Biochemistry Epitope Cell Line Peptide Library Drug Resistance Multiple Bacterial Consensus sequence medicine Animals Amino Acid Sequence Cloning Molecular Molecular Biology Peptide sequence Linear epitope Immunochemistry Flow Cytometry Molecular biology Epitope mapping Multidrug Resistance-Associated Proteins Epitope Mapping |
| Zdroj: | Biological Chemistry. 384:139-142 |
| ISSN: | 1431-6730 |
| DOI: | 10.1515/bc.2003.014 |
| Popis: | In order to study the structure of the multidrug resistance-associated protein (MRP1), which is one of the most important members of the ATP-binding cassette (ABC) protein family acting as drug-efflux systems, we have developed an epitope mapping-based strategy. By means of the mAb MRPr1, we have immunoselected clones from two distinct random peptide libraries displayed on phages and have identified several peptide sequences mimicking the internal conformation of this 190 kDa multidrug transporter protein. Phage clones able to block the immunolabeling of the MRPr1 antibody to MRP1-overexpressing multidrug resistance (MDR) H69/AR cells were isolated and, after sequencing the corresponding inserts, their amino acid sequence was compared to that of MRP1. This analysis led to the identification of the consensus sequence L.SLNWED, corresponding to the MRP1 segment LWSLNKED (residues 241–248). This MRP1sequence is partially overlapping with the MRPr1 epitope GSDL WSLNKE (residues 238–247) previously mapped using peptide scanning techniques. These results demonstrate the high reliability of phage display technology to study not only the topography of complex integral membrane proteins such as MRP1, but also to help identify critical residues participating in the formation of the epitope structure. |
| Databáze: | OpenAIRE |
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