Orai1- and Orai2-, but not Orai3-mediated ICRAC is regulated by intracellular pH.

Grigori Y. Rychkov, Fiona H. Zhou, Melissa K. Adams, Stuart M. Brierley, Linlin Ma, Greg J. Barritt

Research output: Contribution to journalArticlepeer-review

Abstract

Three Orai (Orai1, Orai2 and Orai3) and two STIM (STIM1 and STIM2; stromal interaction molecule) mammalian protein homologues constitute major components of the store-operated Ca2+ entry mechanism. When co-expressed with STIM1, Orai1, Orai2 and Orai3 form highly selective Ca2+ channels with properties of Ca2+ release activated Ca2+ (CRAC) channels. Despite the high level of homology between Orai proteins, CRAC channels formed by different Orai isoforms have distinctive properties, particularly with regards to Ca2+ dependent inactivation, inhibition/potentiation by 2-APB and sensitivity to reactive oxygen species. This study characterises and compares the regulation of Orai1, Orai2- and Orai3-mediated CRAC current (ICRAC) by intracellular pH. Using whole-cell patch clamping of HEK293T cells heterologously expressing Orai and STIM1 we show that ICRAC formed by each Orai homologue has a unique sensitivity to changes in intracellular pH (pHi). Orai1-mediated ICRAC exhibits a strong dependence on pHi of both current amplitude and the kinetics of Ca2+ dependent inactivation. In contrast, Orai2 amplitude, but not kinetics, depends on pHi, whereas Orai3 shows no dependence on pHi at all. Investigation of different Orai1-Orai3 chimeras suggests that pHi dependence of Orai1 resides in both, the N-terminus and intracellular loop 2, and may also involve pH- dependent interactions with STIM1.

Original languageEnglish
JournalUnknown Journal
DOIs
Publication statusPublished or Issued - 2 Nov 2020

Keywords

  • Ca dependent inactivation
  • Gating
  • ICRAC
  • Orai3
  • PH
  • STIM1

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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