Quantification and visualization of phosphoinositides by quantum dot-labeled specific binding-domain probes
Journal of Lipid Research, ISSN: 0022-2275, Vol: 53, Issue: 4, Page: 810-819
2012
- 16Citations
- 50Captures
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Example: if you select the 1-year option for an article published in 2019 and a metric category shows 90%, that means that the article or review is performing better than 90% of the other articles/reviews published in that journal in 2019. If you select the 3-year option for the same article published in 2019 and the metric category shows 90%, that means that the article or review is performing better than 90% of the other articles/reviews published in that journal in 2019, 2018 and 2017.
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Metrics Details
- Citations16
- Citation Indexes16
- 16
- CrossRef12
- Captures50
- Readers50
- 50
Article Description
Phosphoinositides (PI) play important regulatory roles in cell physiology. Localization and quantitation of PIs within the cell is necessary to understand their precise function. Currently, ectopic expression of green fluorescent protein (GFP)-fused PI-binding domains is used to visualize PIs localized to the cell membrane. However, ectopically expressed PI-binding domains may compete with endogenous binding proteins, thus altering the physiological functions of the PIs. Here, we establish a novel method for quantification and visualization of PIs in cells and tissue samples using PI-binding domains labeled with quantum dots (Qdot) as specific probes. This method allowed us to simultaneously quantify three distinct PIs, phosphatidylinositol 3,4,5-triphosphatase [PtdIns(3,4,5)P 3 ), PtdIns(3,4)P 2, and PtdIns(4,5)P 2, in crude acidic lipids extracted from insulin-stimulated cells. In addition, the method allowed the PIs to be visualized within fixed cells and tissues. Sequential and spatial changes in PI production and distribution were detected in platelet-derived growth factor (PDGF)-stimulated NRK49F cells. We also observed accumulation of PtdIns(3,4)P 2 at the dorsal ruffle in PDGF-stimulated NIH3T3 cells. Finally, we found PtdIns(3,4,5)P 3 was enriched in lung cancer tissues, which also showed high levels of phosphorylated Akt. Our new method to quantify and visualize PIs is expected to provide further insight into the role of lipid signaling in a wide range of cellular events.
Bibliographic Details
http://www.sciencedirect.com/science/article/pii/S0022227520405759; http://dx.doi.org/10.1194/jlr.d019547; http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=84859395741&origin=inward; http://www.ncbi.nlm.nih.gov/pubmed/22308508; http://www.jlr.org/lookup/doi/10.1194/jlr.D019547; https://syndication.highwire.org/content/doi/10.1194/jlr.D019547; https://linkinghub.elsevier.com/retrieve/pii/S0022227520405759; https://dx.doi.org/10.1194/jlr.d019547
American Society for Biochemistry & Molecular Biology (ASBMB)
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