Development of nonspecific BRET-based biosensors to monitor plasma membrane inositol lipids in living cells

József T. Tóth, Gergő Gulyás, László Hunyady, Péter Várnai

Research output: Chapter

1 Citation (Scopus)

Abstract

There are several difficulties to face when investigating the role of phosphoinositides. Although they are present in most organelles, their concentration is very low, sometimes undetectable with the available methods; moreover, their level can quickly change upon several external stimuli. Here we introduce a newly improved lipid sensor tool-set based on the balanced expression of luciferase-fused phosphoinositide recognizing protein domains and a Venus protein targeted to the plasma membrane, allowing us to perform Bioluminescence Resonance Energy Transfer (BRET) measurements that reflect phosphoinositide changes in a population of transiently transfected cells. This method is highly sensitive, specific, and capable of semiquantitative characterization of plasma membrane phosphoinositide changes with high temporal resolution.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages23-34
Number of pages12
DOIs
Publication statusPublished - jan. 1 2019

Publication series

NameMethods in Molecular Biology
Volume1949
ISSN (Print)1064-3745

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Tóth, J. T., Gulyás, G., Hunyady, L., & Várnai, P. (2019). Development of nonspecific BRET-based biosensors to monitor plasma membrane inositol lipids in living cells. In Methods in Molecular Biology (pp. 23-34). (Methods in Molecular Biology; Vol. 1949). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9136-5_3