Isolation and characterization of lecithin-cholesterol acyltransferase from hog plasma

Yong B. Park, Andras G. Lacko

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Lecithin-cholesterol acyltransferase (EC 2.3.1.43) was purified from hog plasma by a highly efficient procedure. The final enzyme preparation was purified 30000-fold over the starting material and was homogeneous as indicated by polyacrylamide gel electrophoreses in the presence of both SDS and urea. The purified hog lecithin-cholesterol acyltransferase had an apparent molecular weight of 66000 on SDS-poly-acrylamide gel electrophoresis and HPLC and was found to contain about 21.4% (w/w) carbohydrate-hexose, 11.3%; hexosamine, 1.9%; sialic acid, 8.2%. The amino acid composition analysis showed that hog lecithin-cholesterol acyltransferase contains four half cystines per mol; two cysteines were titrated at neutral pH with 5,5'-dithiobis(2-nitrobenzoic acid). Nearly all the phenolic groups were unavailable to the solvent at neutral pH, while they become exposed at around pH 11. Hog lecithin-cholesterol acyltransferase was found to be associated with HDL in the plasma and it prefers HDL as a substrate. The physicochemical properties of hog lecithin-cholesterol acyltransferase were generally similar to those of the human and the rat enzyme.

Original languageEnglish
Pages (from-to)179-190
Number of pages12
JournalBiochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism
Volume877
Issue number1
DOIs
StatePublished - 11 Jun 1986

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Phosphatidylcholine-Sterol O-Acyltransferase
Plasmas
Cysteine
Dithionitrobenzoic Acid
Hexosamines
Hexoses
N-Acetylneuraminic Acid
Enzymes
Polyacrylates
Electrophoresis
Acrylamide
Urea
Rats
Gels
Molecular weight
Carbohydrates
Amino Acids
Molecular Weight
High Pressure Liquid Chromatography
Substrates

Keywords

  • (Porcine)
  • Amino acid composition
  • Enzyme purification
  • Lecithin-cholesterol acyltransferase

Cite this

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title = "Isolation and characterization of lecithin-cholesterol acyltransferase from hog plasma",
abstract = "Lecithin-cholesterol acyltransferase (EC 2.3.1.43) was purified from hog plasma by a highly efficient procedure. The final enzyme preparation was purified 30000-fold over the starting material and was homogeneous as indicated by polyacrylamide gel electrophoreses in the presence of both SDS and urea. The purified hog lecithin-cholesterol acyltransferase had an apparent molecular weight of 66000 on SDS-poly-acrylamide gel electrophoresis and HPLC and was found to contain about 21.4{\%} (w/w) carbohydrate-hexose, 11.3{\%}; hexosamine, 1.9{\%}; sialic acid, 8.2{\%}. The amino acid composition analysis showed that hog lecithin-cholesterol acyltransferase contains four half cystines per mol; two cysteines were titrated at neutral pH with 5,5'-dithiobis(2-nitrobenzoic acid). Nearly all the phenolic groups were unavailable to the solvent at neutral pH, while they become exposed at around pH 11. Hog lecithin-cholesterol acyltransferase was found to be associated with HDL in the plasma and it prefers HDL as a substrate. The physicochemical properties of hog lecithin-cholesterol acyltransferase were generally similar to those of the human and the rat enzyme.",
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Isolation and characterization of lecithin-cholesterol acyltransferase from hog plasma. / Park, Yong B.; Lacko, Andras G.

In: Biochimica et Biophysica Acta (BBA)/Lipids and Lipid Metabolism, Vol. 877, No. 1, 11.06.1986, p. 179-190.

Research output: Contribution to journalArticle

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AB - Lecithin-cholesterol acyltransferase (EC 2.3.1.43) was purified from hog plasma by a highly efficient procedure. The final enzyme preparation was purified 30000-fold over the starting material and was homogeneous as indicated by polyacrylamide gel electrophoreses in the presence of both SDS and urea. The purified hog lecithin-cholesterol acyltransferase had an apparent molecular weight of 66000 on SDS-poly-acrylamide gel electrophoresis and HPLC and was found to contain about 21.4% (w/w) carbohydrate-hexose, 11.3%; hexosamine, 1.9%; sialic acid, 8.2%. The amino acid composition analysis showed that hog lecithin-cholesterol acyltransferase contains four half cystines per mol; two cysteines were titrated at neutral pH with 5,5'-dithiobis(2-nitrobenzoic acid). Nearly all the phenolic groups were unavailable to the solvent at neutral pH, while they become exposed at around pH 11. Hog lecithin-cholesterol acyltransferase was found to be associated with HDL in the plasma and it prefers HDL as a substrate. The physicochemical properties of hog lecithin-cholesterol acyltransferase were generally similar to those of the human and the rat enzyme.

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