| Fluorescence image screening for chemical compounds modifying cholesterol metabolism and distribution. | |
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MedLine Citation:
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PMID: 21862703 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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An automated fluorescence microscopy assay using a non-toxic cholesterol-binding protein, toxin domain 4 (D4) (Shimada et al. Eur J Biochem 269: 6195 (2002)), was developed, in order to identify chemical compounds modifying intracellular cholesterol metabolism and distribution. Using this method, we screened a library of 1056 compounds and identified 35 compounds that decreased D4-binding to the cell surface. Among them, 8 compounds were already reported to alter the biosynthesis or the intracellular distribution of cholesterol. The remaining 27 hit compounds were further analyzed biochemically and histochemically. Cell staining with another fluorescent cholesterol probe, filipin, revealed that 17 compounds accumulated cholesterol in the late endosomes. Five compounds decreased cholesterol biosynthesis and two compounds inhibited the binding of D4 to the membrane. This visual screening method based on the cholesterol specific probe D4 in combination with biochemical analyses, is a cell-based sensitive technique to identify new chemical compounds modifying cholesterol distribution and metabolism. Furthermore, it is suitable to high-throughput analysis for drug discovery. |
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Authors:
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Reiko Ishitsuka; Tamio Saito; Hiroyuki Osada; Yoshiko Ohno-Iwashita; Toshihide Kobayashi |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-8-23 |
Journal Detail:
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Title: Journal of lipid research Volume: - ISSN: 0022-2275 ISO Abbreviation: - Publication Date: 2011 Aug |
Date Detail:
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Created Date: 2011-8-24 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0376606 Medline TA: J Lipid Res Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
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RIKEN, Japan; |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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