Selective extraction of lecithin:cholesterol acyltransferase (EC 2.3.1.43) from human plasma.
A method for the rapid extraction of lecithin:cholesterol acyltransferase (LCAT) from human plasma or serum has been developed. The method is based on direct treatment of acidified plasma of fully conserved enzyme activity, with the strong ion exchanger Q-Sepharose, which under the experimental conditions bound all LCAT but only about 10% of the total protein content of the plasma, no albumin and essentially no lipoproteins. This corresponds to a 10-fold purification. Only traces of apolipoprotein A-I remained in the quantitatively desorbed LCAT preparation which, however, contained a residual fraction of apolipoprotein D and acidic plasma proteins. The present one-step procedure for extraction of LCAT in high yields from human plasma represents a simple and efficient alternative to the first step in previously described methods for preparation of the enzyme to homogeneity.
Holmquist
Leif
L
National Institute for Working Life, Ekelundsvägen 16, Solna, S-11279 Stockholm, Sweden. leif.holmquist@niwl.se
D000838:Anions; D016632:Apolipoprotein A-I / Q000737:chemistry; D001053:Apolipoproteins / Q000737:chemistry; D053399:Apolipoproteins D; D002852:Chromatography, Ion Exchange; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008074:Lipoproteins; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000097:blood* / Q000302:isolation & purification*; D012685:Sepharose / Q000737:chemistry
D000838:Anions; D016632:Apolipoprotein A-I; D001053:Apolipoproteins; D053399:Apolipoproteins D; D008074:Lipoproteins; D012685:Sepharose; D007862:Phosphatidylcholine-Sterol O-Acyltransferase