caa a22 4500 378849670 CHVBK 20180305123320.0 cr unu---uuuuu 161128e20030127xx s 000 0 eng 10.1515/BC.2003.014 doi (NATIONALLICENCE)gruyter-10.1515/BC.2003.014 Identification by Phage Display of the LinearContinuous MRPr1 Epitope in the MultidrugResistance-Associated Protein (MRP1) [Elektronische Daten] [M. Flego, V. Mennella, F. Moretti, F. Poloni, M.L. Dupuis, A. Ascione, S. Barca, F. Felici, M. Cianfriglia] 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. Copyright © 2003 by Walter de Gruyter GmbH & Co. KG Biochemistry nationallicence Molecular biology nationallicence Cellular biology nationallicence Flego M. aut Mennella V. aut Moretti F. aut Poloni F. aut Dupuis M.L. aut Ascione A. aut Barca S. aut Felici F. aut Cianfriglia M. aut Biological Chemistry Walter de Gruyter 384/1(2003-01-27), 139-142 1431-6730 384:1<139 2003 384 bchm https://doi.org/10.1515/BC.2003.014 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/BC.2003.014 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Flego M. aut NATIONALLICENCE 700 1- Mennella V. aut NATIONALLICENCE 700 1- Moretti F. aut NATIONALLICENCE 700 1- Poloni F. aut NATIONALLICENCE 700 1- Dupuis M.L. aut NATIONALLICENCE 700 1- Ascione A. aut NATIONALLICENCE 700 1- Barca S. aut NATIONALLICENCE 700 1- Felici F. aut NATIONALLICENCE 700 1- Cianfriglia M. aut NATIONALLICENCE 773 0- Biological Chemistry Walter de Gruyter 384/1(2003-01-27), 139-142 1431-6730 384:1<139 2003 384 bchm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter