Rapid fluorometric assay of LDL receptor activity by DiI-labeled LDL.

Stephan, Zouhair F.; Yurachek, E.C.

doi:10.1016/s0022-2275(20)40759-x

Cited by 195 publications

(32 citation statements)

References 22 publications

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“…Binding and cell association of lipoproteins to cells were carried using lipoproteins labeled with the fluorescence probe DiI, a technique extensively used (33,48,49). Cell association of DiI-labeled lipoproteins to COS-7 cells was performed by incubating 5 ϫ 10 4 cells grown on coverslips with 5 g/ml of DiI-labeled lipoproteins in PBS containing 1 m m CaCl 2 , 1 m m MgCl 2 for 1 h at 37 Њ C. Binding of DiI-labeled lipoproteins to Sf9 cells was carried out by incubating cells in 100-200 l of PBS containing 1 m m CaCl 2 , 1 m m MgCl 2 with DiIlipoproteins at 5 g/ml for 2 h at 4 Њ C or 1 h at room temperature.…”

Section: Lipoprotein Binding Assaysmentioning

confidence: 99%

Human CD36 is a high affinity receptor for the native lipoproteins HDL, LDL, and VLDL

Calvo

Gómez-Coronado

Suárez

et al. 1998

Journal of Lipid Research

330

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Mouse and hamster SR-BI glycoproteins and their putative human counterpart CLA-I are so far the only scavenger receptors known to bind both native and modified lipoproteins. CD36, a multigland glycoprotein structurally related to SR-BI and CLA-1, has been reported to bind oxidized low density lipoprotein (OxLDL) and acetylated LDL (AcLDL). In this report, we have studied the ability of CD36 to bind native lipoproteins. By transient expression of human CD36 in mammalian and insect cells, we demonstrate that CD36 is a high affinity receptor for the native lipoproteins HDL, LDL, VLDL, and, as previously reported, for OxLDL and AcLDL. The specificity of these interactions is supported by the dosedependent inhibitory effect of a monoclonal antibody against CD36. Furthermore, at least for HDL, binding to CD36 does not require the presence of apoE. These findings, together with preferential expression of CD36 in tissues performing very active fatty acid metabolism (skeletal muscle, heart, mammary epithelium, and adipose tissue) and its involvement in foam cell formation (macrophages), suggest that binding of lipoproteins to CD36 might contribute to the regulation of lipid metabolism, and to the pathogenesis of atherosclerosis. -

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Section: Lipoprotein Binding Assaysmentioning

confidence: 99%

Human CD36 is a high affinity receptor for the native lipoproteins HDL, LDL, and VLDL

Calvo

Gómez-Coronado

Suárez

et al. 1998

Journal of Lipid Research

330

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“…We first tested whether the EDEM3 gene deletion might affect the VLDL uptake. The KO and control Huh7 and HepG2 clones were incubated with human Dil-VLDL (Stephan and Yurachek, 1993). We found that EDEM3 deficiency strongly increased the VLDL uptake in both Huh7 (by 1.6 times) and HepG2 (by~5.2 times) cells (Figure 3).…”

Section: Deletion Of Edem3 Gene Strongly Increased Vldl But Not Ldl Umentioning

confidence: 88%

“…The amount of ApoB-100 was normalized with cell numbers. For VLDL/LDL uptake assays, EDEM3 KO and control HepG2 and Huh7 cells with, and the KO cells with WT/mutant EDEM3 complementation and relevant controls were incubated in serum-free medium (2 hrs) followed by incubation in serum-free medium containing Dil-VLDL and Dil-LDL (Stephan and Yurachek, 1993) (5 µg/ml, Invitrogen) for 1 hr. The cells were then fixed and analyzed under confocal microscope as described below.…”

Section: Declaration Of Interestsmentioning

confidence: 99%

EDEM3 Modulates Plasma Triglyceride Level through Its Regulation of LRP1 Expression

Peloso

Nagai

et al. 2020

iScience

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“…The effect of the scFv-anti-LDL(-)-MCMN-Zn nanoformulation on the uptake of LDL(-) by both species of macrophages was evaluated by flow cytometry. Cells were treated with 10 4 particles/mL of scFv-anti-LDL(-)-MCMN-Zn nanoformulation (containing 6.25 ug of scFv protein/ml of nanoformulation) and LDL(-) solution (containing 37.5 µg of LDL(-) protein/mL) individually or combined for 3 h at 37 • C. The LDL(-) was labeled with the fluorophore DiI(C 18 ) (1,1 ′ -Dioctadecyl-3,3,3 ′ ,3 ′ -Tetramethylindocarbocyanine Perchlorate, #D282, Life Technologies, USA) (16).…”

Section: Cellular Uptake Of Ldl(-)mentioning

confidence: 99%

scFv-Anti-LDL(-)-Metal-Complex Multi-Wall Functionalized-Nanocapsules as a Promising Tool for the Prevention of Atherosclerosis Progression

et al. 2021

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Atherosclerosis can be originated from the accumulation of modified cholesterol-rich lipoproteins in the arterial wall. The electronegative LDL, LDL(-), plays an important role in the pathogenesis of atherosclerosis once this cholesterol-rich lipoprotein can be internalized by macrophages, contributing to the formation of foam cells, and provoking an immune-inflammatory response. Herein, we engineered a nanoformulation containing highly pure surface-functionalized nanocapsules using a single-chain fragment variable (scFv) reactive to LDL(-) as a ligand and assessed whether it can affect the LDL(-) uptake by primary macrophages and the progression of atherosclerotic lesions in Ldlr−/− mice. The engineered and optimized scFv-anti-LDL(-)-MCMN-Zn nanoformulation is internalized by human and murine macrophages in vitro by different endocytosis mechanisms. Moreover, macrophages exhibited lower LDL(-) uptake and reduced mRNA and protein levels of IL1B and MCP1 induced by LDL(-) when treated with this new nanoformulation. In a mouse model of atherosclerosis employing Ldlr−/− mice, intravenous administration of scFv-anti-LDL(-)-MCMN-Zn nanoformulation inhibited atherosclerosis progression without affecting vascular permeability or inducing leukocytes-endothelium interactions. Together, these findings suggest that a scFv-anti-LDL(-)-MCMN-Zn nanoformulation holds promise to be used in future preventive and therapeutic strategies for atherosclerosis.

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Rapid fluorometric assay of LDL receptor activity by DiI-labeled LDL.

Cited by 195 publications

References 22 publications

Human CD36 is a high affinity receptor for the native lipoproteins HDL, LDL, and VLDL

Human CD36 is a high affinity receptor for the native lipoproteins HDL, LDL, and VLDL

EDEM3 Modulates Plasma Triglyceride Level through Its Regulation of LRP1 Expression

scFv-Anti-LDL(-)-Metal-Complex Multi-Wall Functionalized-Nanocapsules as a Promising Tool for the Prevention of Atherosclerosis Progression

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