Exploiting Cameleon Probes to Investigate Organelles Ca2+ Handling

Methods Mol Biol. 2019:1925:15-30. doi: 10.1007/978-1-4939-9018-4_2.

Abstract

Calcium ion (Ca2+) is a ubiquitous intracellular messenger able to generate versatile intracellular signals that modulate a large variety of functions in virtually every cell type. Chemical and genetic biosensors, targeted to different subcellular compartments, have been developed and continuously improved to monitor Ca2+ dynamics in living cells. Here we describe the usage of Förster resonance energy transfer (FRET)-based Cameleon probes to investigate Ca2+ influx across the plasma membrane (PM) or Ca2+ release from the main intracellular Ca2+ store, the endoplasmic reticulum (ER).

Keywords: Ca2+ imaging; Ca2+ signaling; Cameleon; Endoplasmic reticulum; FRET; IP3 receptor; SOCE.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / metabolism*
  • Calcium / analysis
  • Calcium / metabolism*
  • Calcium Signaling
  • Cations, Divalent / analysis
  • Cations, Divalent / metabolism
  • Cell Line
  • Cell Membrane / metabolism*
  • Endoplasmic Reticulum / metabolism*
  • Fluorescence Resonance Energy Transfer / methods*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / metabolism*
  • HeLa Cells
  • Humans
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / metabolism*
  • Optical Imaging / methods
  • Peptides / chemistry
  • Peptides / metabolism
  • Protein Binding
  • Protein Conformation

Substances

  • Bacterial Proteins
  • Cations, Divalent
  • Fluorescent Dyes
  • Luminescent Proteins
  • Peptides
  • yellow fluorescent protein, Bacteria
  • Green Fluorescent Proteins
  • Calcium