Purification and characterization of a phosphatidylinositol transfer protein from human platelets.
We report the purification of a phospholipid transfer protein from human platelets. This protein preferentially transfers phosphatidylinositol, with phosphatidylcholine and phosphatidylglycerol being transferred to a lesser extent. Phosphatidylethanolamine is not transferred. Transfer activity is detected by measuring the transfer of radiolabeled phospholipids between two populations of small unilamellar vesicles. The protein was purified approximately 1000-fold over the platelet cytosol by chromatography on Sephadex G-75, sulfooxyethyl cellulose, and hydroxylapatite. The molecular weight of this protein appears to be 28 000 as determined by gel filtration chromatography. When the purified protein is analyzed on sodium dodecyl sulfate-polyacrylamide gels, two major components and several minor ones are observed. The molecular weight of the two major bands are 28 600 and 29 200. Isoelectric focusing of the platelet cytosol yielded phosphatidylinositol and phosphatidylcholine transfer activity at pH 5.6 and 5.9. The platelet phospholipid transfer protein is able to catalyze the transfer of phosphatidylinositol and phosphatidylcholine between vesicles and human platelet plasma membranes. One possible physiological role for this transfer protein is an involvement in the rapid turnover of inositol-containing lipids which occurs upon exposure of platelets to various stimuli.
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D001792:Blood Platelets / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification*; D002384:Catalysis; D002462:Cell Membrane / Q000378:metabolism; D002482:Cellulose / Q000031:analogs & derivatives; D055598:Chemical Phenomena; D002621:Chemistry; D002845:Chromatography / Q000379:methods; D003600:Cytosol / Q000378:metabolism; D003911:Dextrans; D017886:Durapatite; D006801:Humans; D006882:Hydroxyapatites; D007525:Isoelectric Focusing; D008565:Membrane Proteins; D008970:Molecular Weight; D049294:Phospholipid Transfer Proteins; D029701:Saccharomyces cerevisiae Proteins
D002352:Carrier Proteins; D003911:Dextrans; D006882:Hydroxyapatites; D008565:Membrane Proteins; D049294:Phospholipid Transfer Proteins; C118289:SEC24 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D002482:Cellulose; C025614:sephadex; C046567:sulfoxyethyl cellulose; D017886:Durapatite