Details

Autor(en) / Beteiligte

• Bassé, Fran¸ois
• Stout, James G.
• Sims, Peter J.
• Wiedmer, Therese

Titel

Isolation of an Erythrocyte Membrane Protein that Mediates Ca2+-dependent Transbilayer Movement of Phospholipid

Ist Teil von

• The Journal of biological chemistry, 1996-07, Vol.271 (29), p.17205-17210

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

1996

Quelle

MEDLINE

Beschreibungen/Notizen

• Elevation of intracellular Ca2+ in erythrocytes, platelets, and other cells initiates rapid redistribution of plasma membrane phospholipids (PL) between inner and outer leaflets, collapsing the normal asymmetric distribution. Consequently, phosphatidylserine and other lipids normally sequestered to the inner leaflet become exposed at the cell surface. This Ca2+-induced mobilization of phosphatidylserine to the surface of activated, injured, or apoptotic cells confers a procoagulant property to the plasma membrane, which promotes fibrin clotting and provides a signal for cell removal by the reticuloendothelial system. To identify the constituent of the membrane that mediates this Ca2+-dependent “PL scramblase” activity, we undertook purification and reconstitution of membrane component(s) with this activity from detergent extracts of erythrocyte ghosts depleted of cytoskeleton. Active fractions were identified by their capacity to mediate the Ca2+-dependent redistribution of 7-nitrobenz-2-oxa-1,3-diazol-4-yl-labeled PL between leaflets of reconstituted proteoliposomes. This PL scramblase activity co-eluted through multiple chromatographic steps with a single polypeptide of ∼37 kDa, which was purified to apparent homogeneity as resolved by silver staining. The activity associated with this protein band was inactivated by trypsin. The isolated protein reconstituted in proteoliposomes mediated nonselective, bidirectional transport of 7-nitrobenz-2-oxa-1,3-diazol-4-yl-PL between membrane leaflets, with half-maximal activation between 20 and 60 µM Ca2+ (saturation >100 µM), mimicking the Ca2+-dependent transbilayer lipid movement intrinsic to the erythrocyte membrane.