Interaction of the P-glycoprotein multidrug transporter (MDR1) with high affinity peptide chemosensitizers in isolated membranes, reconstituted systems, and intact cells

Frances J. Sharom, Xiaohong Yu, Peihua Lu, Ronghua Liu, Joseph W K Chu, Katalin Szabó, M. Müller, Curtis D. Hose, Anne Monks, A. Váradi, J. Seprődi, B. Sarkadi

Research output: Contribution to journalArticle

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Abstract

P-Glycoprotein-mediated multidrug resistance can be reversed by the action of a group of compounds known as chemosensitizers. The interactions with P-glycoprotein of two novel hydrophobic peptide chemosensitizers (reversins 121 and 205) have been studied in model systems in vitro, and in a variety of MDR1-expressing intact tumor cells. The reversins bound to purified P-glycoprotein with high affinity (77-154 nM), as assessed by a quenching assay using fluorescently labeled purified protein. The peptides modulated P-glycoprotein ATPase activity in Sf9 insect cell membranes expressing human MDR1, plasma membrane vesicles from multidrug-resistant cells, and reconstituted proteoliposomes. Both peptides induced a large stimulation of ATPase activity; however, higher concentrations, especially of reversin 205, led to inhibition. This pattern was different from that of simple linear peptides, and resembled that of chemosensitizers such as verapamil. In both membrane vesicles and reconstituted proteoliposomes, 1-2 μM reversins were more effective than cyclosporin A at blocking colchicine transport. Reversin 121 and reversin 205 restored the uptake of [3H]daunorubicin and rhodamine 123 in MDR1-expressing cells to the level observed in the drug-sensitive parent cell lines, and also effectively inhibited the extrusion of calcein acetoxymethyl ester from intact cells. In cytotoxicity assays, reversin 121 and reversin 205 eliminated the resistance of MDR1-expressing tumor cells against MDR1-substrate anticancer drugs, and they had no toxic effects in MDR1-negative control cells. We suggest that peptides of the reversin type interact with the MDR1 protein with high affinity and specificity, and thus they may be good candidates for the development of MDR1-modulating agents to sensitize drug resistance in cancer. Copyright (C) 1999 Elsevier Science Inc.

Original languageEnglish
Pages (from-to)571-586
Number of pages16
JournalBiochemical Pharmacology
Volume58
Issue number4
DOIs
Publication statusPublished - Aug 15 1999

Fingerprint

P-Glycoprotein
Membranes
Peptides
Cells
Cell membranes
Adenosine Triphosphatases
Tumors
Assays
Pharmaceutical Preparations
Rhodamine 123
Cell Membrane
Daunorubicin
Poisons
Colchicine
Sf9 Cells
Neoplasms
Cytotoxicity
Verapamil
Cyclosporine
Multiple Drug Resistance

Keywords

  • ATPase activity
  • Azidopine photolabeling
  • Daunorubicin transport
  • Drug binding
  • Drug transport
  • Fluorescent dye transport
  • Peptide derivatives
  • Proteoliposomes

ASJC Scopus subject areas

  • Pharmacology

Cite this

Interaction of the P-glycoprotein multidrug transporter (MDR1) with high affinity peptide chemosensitizers in isolated membranes, reconstituted systems, and intact cells. / Sharom, Frances J.; Yu, Xiaohong; Lu, Peihua; Liu, Ronghua; Chu, Joseph W K; Szabó, Katalin; Müller, M.; Hose, Curtis D.; Monks, Anne; Váradi, A.; Seprődi, J.; Sarkadi, B.

In: Biochemical Pharmacology, Vol. 58, No. 4, 15.08.1999, p. 571-586.

Research output: Contribution to journalArticle

Sharom, Frances J. ; Yu, Xiaohong ; Lu, Peihua ; Liu, Ronghua ; Chu, Joseph W K ; Szabó, Katalin ; Müller, M. ; Hose, Curtis D. ; Monks, Anne ; Váradi, A. ; Seprődi, J. ; Sarkadi, B. / Interaction of the P-glycoprotein multidrug transporter (MDR1) with high affinity peptide chemosensitizers in isolated membranes, reconstituted systems, and intact cells. In: Biochemical Pharmacology. 1999 ; Vol. 58, No. 4. pp. 571-586.
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