Abstract
Phosphatidylinositol‐4,5‐bisphosphate (PtdIns(4,5)P2), which is a minor component of the plasma membrane, plays important roles in many cellular processes, including signal transduction, exocytosis, endocytosis, and ion channel activation. A method for quantitatively monitoring the PtdIns(4,5)P2 concentration and dynamics in living cells is lacking at present. We have developed a new method of quantifying the PtdIns(4,5)P2 concentration in living cells using a specific PtdIns(4,5)P2 sensor made of the engineered ENTH domain of epsin1 labeled with an environment‐sensitive fluorophore. The PtdIns(4,5)P2 sensor allows robust quantitative determination of spatiotemporal fluctuation of PtdIns(4,5)P2 in giant unilamellar vesicles and mammalian cells by ratiometric analysis. The results demonstrate the spatially heterogeneous distribution of PtdIns(4,5)P2 in the plasma membrane and provide new insight into the fluctuation and dynamics of cellular PtdIns(4,5)P2 in response to various stimuli. Furthermore, this strategy is generally applicable to in situ quantification of other membrane lipids.Supported by NIH GM 68849 and GM76581
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
