Colocalization of Rh polypeptides and the aminophospholipid transporter in dilauroylphosphatidylcholine-induced erythrocyte vesicles

Elizabeth M. Bruckheimer, Karen D. Gillum, Alan J. Schroit

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Cytoskeleton-free vesicles released from human red blood cells (RBC) transport exogenously supplied aminophospholipid analogues from the vesicle's outer to inner leaflet at rates comparable to those of normal RBC (Beleznay et al. (1993) Biochemistry 32, 3146-3152). Because polypeptides associated with the Rh blood group system have been implicated in the transbilayer movement of phosphatidylserine (PS), we investigated the relationship and co-localization of the aminophospholipid translocase and Rh in dilauroylphosphatidylcholine-inducted RBC vesicles. The transbilayer movement of fluorescent (NBD-PS) and photoactivatable (125I-N3-PS) PS in RBC vesicles was ATI'- and temperature-dependent. Inhibition of PS transport by sulfhydryl reagents could be accomplished by direct vesicle treatment or by treating RBC before vesiculation. In the case of diamide- and pyridyldithioethylamine-mediated inhibition, NBD-PS transport could be restored by reduction with dithiothreitol, indicating that the movement of the PS transporter into the emerging vesicle was independent of the oxidative status of membrane sulfhydryls. The presence of Rh polypeptides in the vesicles was verified by direct immunoprecipitation of isotopically-labeled Rh and semi-quantified by antibody adsorption assays. Similar to the movement of the PS transporter, localization of Rh polypeptides in the vesicle membrane was independent of the red cell's oxidative status. These results show that the PS translocase and Rh-related proteins colocalize in RBC vesicles suggesting that these proteins may be members of a multicomponent complex that plays a role in lipid movement and the generation of membrane lipid asymmetry.

Original languageEnglish (US)
Pages (from-to)147-154
Number of pages8
JournalBBA - Biomembranes
Volume1235
Issue number1
DOIs
StatePublished - Apr 12 1995

Keywords

  • Lipid transporter
  • Phospholipid
  • Vesicle

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Cell Biology

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