IDENTIFICATION OF PROTEINS ASSOCIATED WITH APOLIPOPROTEIN A-I-CONTAINING LIPOPROTEINS PURIFIED BY SELECTED-AFFINITY IMMUNOSORPTION

被引:52
作者
KUNITAKE, ST [1 ]
CARILLI, CT [1 ]
LAU, K [1 ]
PROTTER, AA [1 ]
NAYAVIGNE, J [1 ]
KANE, JP [1 ]
机构
[1] SCIOS INC, Mountain View, CA 94043 USA
关键词
D O I
10.1021/bi00174a003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The isolation of apolipoprotein A-I-containing lipoproteins [Lp(A-I)] by selected-affinity immunosorption minimizes the loss of associated proteins that occurs during the isolation of high-density lipoproteins (HDL) by sequential ultracentrifugation. We have used two-dimensional gel electrophoretic analysis to separate the proteins associated with Lp(A-I). Using a combination of amino acid sequencing of transblotted proteins and Western blotting with specific antisera, we have identified a number of associated proteins. The positions of the apolipoproteins (ape) A-I, A-II, A-IV, C-III, D, and E were located on the gels. Lecithin-cholesterol acyltransferase and cholesteryl ester transfer protein were identified in association with Lp(A-I) to a greater extent than found associated with HDL after centrifugation. In addition to those proteins previously identified in association with HDL, we detected a number of plasma proteins associated with Lp(A-I), namely, fibrinogen, haptoglobin, proline-rich protein (C4b-binding protein), and apolipoprotein J (SP40,40 sulfated grycoprotein). The coisolation of these proteins with Lp(A-I) does not appear to be an artifact in that they have very low affinity for a sham column containing covalently bound preimmune goat IgG in place of the anti-apoA-I IgG. These findings suggest that in addition to apolipoproteins that exist largely in association with lipoproteins there is another class of proteins which exist both in lipoprotein- associated form and in the dispersed state. Detection and identification of these lipoprotein-associated proteins may aid in the mechanistic determination of a humber of observed functions attributed to HDL.
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页码:1988 / 1993
页数:6
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