P-glycoprotein retains function when reconstituted into a sphingolipid- and cholesterol-rich environment

Szabolcs Modok, Catherine Heyward, Richard Callaghan

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

P-glycoprotein (P-gp) appears to be associated within specialized raftlike membrane microdomains. The activity of P-gp is sensitive to its lipid environment, and a functional association hi raft microdomains will require that P-gp retains activity in the microenvironment. Purified hamster P-gp was reconstituted in liposomes comprising sphingomyelin and cholesterol, both highly enriched in membrane microdomains and known to impart a liquid-ordered phase to bilayers. The activity of P-gp was compared with that of proteoliposomes composed of crude egg phosphatidylcholine (unsaturated) or dipalmitoyl phosphatidylcholine (saturated) in the presence or absence of cholesterol. The maximal rate of ATP hydrolysis was not significantly altered by the nature of the lipid species. However, the potencies of nicardipine and XR9576 to modulate the ATPase activity of P-gp were increased in the sphingolipid-based proteoliposomes. The drug-P-gp interaction was investigated by measurement of the rates of [3H]XR9576 association and dissociation from the transporter. The lipid environment of P-gp did not affect these kinetic parameters of drug binding. In summary, P-gp retains function in liquid-ordered cholesterol and sphingolipid model membranes in which the communication between the transmembrane and the nucleotide binding domains after drug binding to the protein is more efficient.

Original languageEnglish
Pages (from-to)1910-1918
Number of pages9
JournalJournal of Lipid Research
Volume45
Issue number10
DOIs
Publication statusPublished - Oct 1 2004

    Fingerprint

Keywords

  • ATP binding cassette transporter
  • Drug binding
  • Lipid raft
  • Liquid-ordered phase
  • Membrane microdomain

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this