Level of expression of phospholipid scramblase regulates induced movement of phosphatidylserine to the cell surface.
We recently identified a 35-kDa erythrocyte membrane protein, phospholipid scramblase, that promotes Ca2+-dependent transbilayer movement of phosphatidylserine (PS) and other phospholipids (PL) in reconstituted proteoliposomes (Zhou, Q., Zhao, J., Stout, J. G., Luhm, R. A., Wiedmer, T., and Sims, P. J. (1997) J. Biol. Chem. 272, 18240-18244). To determine whether this same protein is responsible for the rapid movement of PS from inner-to-outer plasma membrane leaflets in other cells exposed to elevated cytosolic calcium concentration ([Ca2+]c), we analyzed how induced movement of PS to the cell surface related to expression of PL scramblase. Exposure to Ca2+ ionophore A23187 resulted in rapid PS exposure in those cell lines constitutively high in PL scramblase (HEL, Epstein-Barr virus-transformed B-lymphocytes, and Jurkat), whereas this response was markedly attenuated in cells expressing low amounts of this protein (Raji, HL60, and Dami). To confirm this apparent correlation between PL scramblase expression and PS egress at elevated [Ca2+]c, Raji cells were transfected with PL scramblase cDNA in pEGFP-C2, and stable transformants expressing various amounts of GFP-PL scramblase fusion protein were obtained. Clones expressing GFP-PL scramblase showed distinctly plasma membrane-localized fluorescence. When compared either with untransfected Raji cells or with transformants expressing GFP alone, clones expressing GFP-PL scramblase fusion protein showed increased exposure of PS at the cell surface in response to elevated [Ca2+]c, accompanied by increased expression of membrane catalytic function for the prothrombinase enzyme complex. These data indicate that transfection with PL scramblase cDNA promotes movement of PS to cell surfaces and suggest that this protein normally mediates redistribution of plasma membrane phospholipids in activated, injured, or apoptotic cells.
Zhao
J
J
Blood Center of Southeastern Wisconsin, Milwaukee, Wisconsin 53201, USA.
Zhou
Q
Q
Wiedmer
T
T
Sims
P J
PJ
D001692:Biological Transport; D000001:Calcimycin / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D008565:Membrane Proteins; D010718:Phosphatidylserines; D049294:Phospholipid Transfer Proteins; D000001:Calcimycin