Polyacrylamide gradient gel electrophoresis as a method to measure transfers of radiolabeled cholesteryl esters between several plasma lipoprotein fractions.
In human plasma, cholesteryl esters can redistribute between various lipoprotein fractions through an exchange reaction mediated by the cholesteryl ester transfer protein. In the present study, liquid scintillation counting of polyacrylamide gradient gel fragments was applied to the determination of transfers of radio-labeled cholesteryl esters between different lipoprotein fractions. This method consisted in four successive steps: (1) incubation of total human plasmas with a tracer dose of high-density lipoproteins (HDL) containing tritium 3H-labeled cholesteryl esters, (2) separation of plasma lipoprotein fractions by electrophoresis in 20-160 g/liter polyacrylamide gels, (3) cutting off and dissolution with a NaOCl solution of polyacrylamide gel fragments which contained the different lipoprotein fractions, (4) liquid scintillation counting of the dissolved gel fragments. The present method provided a convenient and accurate method of determining the rate of cholesteryl ester transferred between several plasma lipoprotein classes. As an example, it was applied to the measurement of the rates of radiolabeled cholesteryl esters transferred from either HDL3 or HDL2 toward the two other main plasma lipoprotein fractions, very-low-density lipoproteins and low-density lipoproteins.
Florentin
E
E
Laboratoire de Biochimie des Lipoprotéines, Faculté de Médecine, Dijon, France.
Lagrost
L
L
Lallemant
C
C
Gambert
P
P
D002788:Cholesterol Esters / Q000097:blood*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006801:Humans; D008074:Lipoproteins / Q000097:blood* / Q000302:isolation & purification; D008075:Lipoproteins, HDL / Q000097:blood; D008077:Lipoproteins, LDL / Q000097:blood; D008079:Lipoproteins, VLDL / Q000097:blood; D014316:Tritium
D002788:Cholesterol Esters; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL; D014316:Tritium