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Separation of a plasma phospho lipid transfer protein from cholesterol ester phospho lipid exchange protein

Separation of a plasma phospho lipid transfer protein from cholesterol ester phospho lipid exchange protein

Journal of Biological Chemistry 258(4): 2174-2180

The d [density] > 1.21 g/ml fraction of human plasma stimulates mass transfer of phosphatidylcholine (PC) from egg PC vesicles into plasma high density lipoproteins (HDL) and also exchange of PC and cholesterol esters (CE) between HDL [high density lipoprotein] and low density lipoproteins (LDL). By measuring the facilitated transfer of PC from egg PC vesicles into HDL and the facilitated exchange of cholesterol esters between HDL and LDL, PC transfer and CE exchange activities were monitored through several purification steps. After application of the d > 1.19 fraction to phenyl-Sepharose, both activities were eluted with H2O. However, they were subsequently partially separated by a linear NaCl gradient on a CM-cellulose column. After further chromatography on diethylaminoethylcellulose, fractions showing highest PC transfer or CE exchange activities showed, respectively, no CE exchange or PC transfer activities. Further purification of the PC transfer activity was achieved by adsorption from active ion exchange fractions to PC vesicles, followed by isolation of vesicles by agarose chromatography and ultracentrifugal flotation. Sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis showed that the Ce exchange and PC transfer activities were associated with proteins of apparent MW = 63,000 and 41,000, respectively. In further experiments, double-labeled HDL ([14C]PC, [3H]CE) were used to measure PC and CE exchange between HDL and LDL. In contrast to the results obtained in the PC transfer assay, the PC and CE exchange activities showed identical distribution through the ion exchange steps. The mass transfer of PC into HDL may be facilitated by a different plasma protein to that enhancing exchange of CE and PC between LDL and HDL.

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Accession: 006391246

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PMID: 6822552

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