Purification and characterization of human plasma lecithin: cholesterol acyltransferase

Abstract
A highly purified (approximately 12,000-fold) homogeneous preparation of human plasma lecithin:cholesterol acyltransferase (EC 2.3.1.43) with 16% yield was obtained by a combination of density ultracentrifugation, high density lipoprotein affinity column chromatography, hydroxylapatite chromatography and finally chromatography on anti-apolipoprotein D immunoglobulin-Sepharose columns to remove apolipoprotein D. This enzyme preparation was homogeneous by the following criteria: a single band by polyacrylamide gel electrophoresis in 8 M urea, a single band on sodium dodecyl sulfate gel electrophoresis with an apparent molecular weight of 68,000 .+-. 1600 and a single protein peak with a molecular weight of 70,000 on a calibrated Sephadex G-100 column. Its amino acid composition was different from human serum albumin and all other apoproteins isolated from lipoprotein fractions.

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