Patch-Clamp Recording of the CRAC Channel Current in STIM-Orai Overexpressing Cells

Methods Mol Biol. 2018:1843:1-16. doi: 10.1007/978-1-4939-8704-7_1.

Abstract

Induced by the depletion of ER calcium store, the calcium influx through calcium release-activated calcium (CRAC) channels is an ubiquitous way of Ca2+ influx for most cell types. This process is mediated by STIM protein, ER calcium sensor, and PM localized Orai calcium channels. Biophysical characterization of this STIM-Orai-mediated current, or ICRAC, with whole-cell patch-clamp technique is essential for revealing the molecular mechanisms underlying the process of STIM-Orai activation or modulation. Here we describe one commonly used procedure of monitoring CRAC activity with whole-cell patch-clamp technique.

Keywords: Calcium release-activated calcium (CRAC) channel; ICRAC; Store-operated Ca2+ entry (SOCE); Whole-cell patch clamp.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Calcium Release Activated Calcium Channels / physiology*
  • Cell Line
  • Data Analysis
  • Endoplasmic Reticulum / metabolism
  • Gene Expression*
  • Humans
  • Ion Channel Gating*
  • Patch-Clamp Techniques* / instrumentation
  • Patch-Clamp Techniques* / methods
  • Software
  • Stromal Interaction Molecules / genetics*
  • Transfection

Substances

  • Calcium Release Activated Calcium Channels
  • Stromal Interaction Molecules