| Reconstituted low density lipoprotein: a vehicle for the delivery of hydrophobic fluorescent probes to cells. | |
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MedLine Citation:
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PMID: 229357 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Previous studies have shown that the cholesteryl ester core of plasma low density lipoprotein (LDL) can be extracted with heptane and replaced with a variety of hydrophobic molecules. In the present report we use this reconstitution technique to incorporate two fluorescent probes, 3-pyrenemethyl-23, 24-dinor-5-cholen-22-oate-3 beta-yl oleate (PMCA oleate) and dioleyl fluorescein, into heptane-extracted LDL. Both fluorescent lipoprotein preparations were shown to be useful probes for visualizing the receptor-mediated endocytosis of LDL in cultured human fibroblasts. When normal fibroblasts were incubated at 37 degrees C with either of the fluorescent LDL preparations, fluorescent granules accumulated in the perinuclear region of the cell. In contrast, fibroblasts from patients with the homozygous form of familial hypercholesterolemia (FH) that lack functional LDL receptors did not accumulate visible fluorescent granules when incubated with the fluorescent reconstituted LDL. A fluorescence-activated cell sorter was used to quantify the fluorescence intensity of individual cells that had been incubated with LDL reconstituted with dioleyl fluorescein. With this technique a population of normal fibroblasts could be distinguished from a population of FH fibroblasts. The current studies demonstrate the feasibility of using fluorescent reconstituted LDL in conjunction with the cell sorter to isolate mutant cells lacking functional LDL receptors. |
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Authors:
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M Krieger; L C Smith; R G Anderson; J L Goldstein; Y J Kao; H J Pownall; A M Gotto; M S Brown |
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Publication Detail:
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Type: Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Journal of supramolecular structure Volume: 10 ISSN: 0091-7419 ISO Abbreviation: J. Supramol. Struct. Publication Date: 1979 |
Date Detail:
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Created Date: 1980-02-15 Completed Date: 1980-02-15 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0330464 Medline TA: J Supramol Struct Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 467-78 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Cells, Cultured Fibroblasts / metabolism Fluorescent Dyes Humans Hypercholesterolemia / blood, genetics Lipoproteins, LDL* / blood Protein Binding Receptors, Drug / metabolism Spectrometry, Fluorescence |
| Chemical | |
Reg. No./Substance:
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0/Fluorescent Dyes; 0/Lipoproteins, LDL; 0/Receptors, Drug |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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