Interaction of Native and Cell-modified Low Density Lipoprotein with Collagen Gel

We have examined the binding of native and cell-modified low density lipoprotein (LDL) to gels of Type I collagen. Diffusion of nativeI-LDL Into the collagen gel was slow, reaching equilibrium after 24 to 48 hours, while L-3H-glucose, a low molecular weight marker, equilibrated in 6 hours. Binding o...

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Veröffentlicht in:Arteriosclerosis, thrombosis, and vascular biology thrombosis, and vascular biology, 1988-09, Vol.8 (5), p.525-534
Hauptverfasser: Hoover, Gayle A, McCormick, Suzanne, Kalant, Norman
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container_end_page 534
container_issue 5
container_start_page 525
container_title Arteriosclerosis, thrombosis, and vascular biology
container_volume 8
creator Hoover, Gayle A
McCormick, Suzanne
Kalant, Norman
description We have examined the binding of native and cell-modified low density lipoprotein (LDL) to gels of Type I collagen. Diffusion of nativeI-LDL Into the collagen gel was slow, reaching equilibrium after 24 to 48 hours, while L-3H-glucose, a low molecular weight marker, equilibrated in 6 hours. Binding ofI-LDL was measured at 48 hours as the amount associated with the collagen after extensive washing. Binding was saturable with an Increasing concentration of LDL. Prior incubation with cell-free culture medium resulted in modest, but progressive, Increases In electrophoretlc mobility and binding to collagen. Incubation with cells produced a marked Increase In electrophoretlc mobility and a 5- to 10-fold Increase In collagen binding; the presence of butyiated hydroxytoluene during Incubation prevented both effects. These changes In LDL were Induced by porcine aortic endothellal cells, smooth muscle cells, human skin flbroblasts, and a variety of cell lines, as well as by acetylatlon. There was a curvilinear relationship between the amount of LDL protein bound and the net negative charge of the LDL; Increasing net charge was associated with progressively greater Increases In binding. These results suggest a potential role for collagen in trapping llpld In the extracellular matrix of arterial Intlma by slowing the diffusion of and by binding LDL. The data also demonstrate that binding of LDL to collagen Is enhanced by modifications that increase its net negative charge.
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Diffusion of nativeI-LDL Into the collagen gel was slow, reaching equilibrium after 24 to 48 hours, while L-3H-glucose, a low molecular weight marker, equilibrated in 6 hours. Binding ofI-LDL was measured at 48 hours as the amount associated with the collagen after extensive washing. Binding was saturable with an Increasing concentration of LDL. Prior incubation with cell-free culture medium resulted in modest, but progressive, Increases In electrophoretlc mobility and binding to collagen. Incubation with cells produced a marked Increase In electrophoretlc mobility and a 5- to 10-fold Increase In collagen binding; the presence of butyiated hydroxytoluene during Incubation prevented both effects. These changes In LDL were Induced by porcine aortic endothellal cells, smooth muscle cells, human skin flbroblasts, and a variety of cell lines, as well as by acetylatlon. There was a curvilinear relationship between the amount of LDL protein bound and the net negative charge of the LDL; Increasing net charge was associated with progressively greater Increases In binding. These results suggest a potential role for collagen in trapping llpld In the extracellular matrix of arterial Intlma by slowing the diffusion of and by binding LDL. 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Diffusion of nativeI-LDL Into the collagen gel was slow, reaching equilibrium after 24 to 48 hours, while L-3H-glucose, a low molecular weight marker, equilibrated in 6 hours. Binding ofI-LDL was measured at 48 hours as the amount associated with the collagen after extensive washing. Binding was saturable with an Increasing concentration of LDL. Prior incubation with cell-free culture medium resulted in modest, but progressive, Increases In electrophoretlc mobility and binding to collagen. Incubation with cells produced a marked Increase In electrophoretlc mobility and a 5- to 10-fold Increase In collagen binding; the presence of butyiated hydroxytoluene during Incubation prevented both effects. These changes In LDL were Induced by porcine aortic endothellal cells, smooth muscle cells, human skin flbroblasts, and a variety of cell lines, as well as by acetylatlon. 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source Alma/SFX Local Collection
title Interaction of Native and Cell-modified Low Density Lipoprotein with Collagen Gel
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