Skip to main content
SpringerLink
Log in

Measurement of the CRAC Channel Fast Ca2+-Dependent Inactivation (FCDI)

  • Protocol
  • First Online:
The CRAC Channel

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1843))

Abstract

Fast Ca2+-dependent inactivation (FCDI) is a safety mechanism limiting Ca2+ entry through some types of Ca2+ channels, including Ca2+ release-activated Ca2+ (CRAC) channels. This type of inactivation is caused by Ca2+, which passes through Ca2+ channel and binds to a specific site within a short distance from the inner mouth of the pore, causing channel to shut.

The main technique that is used to investigate FCDI is whole-cell patch clamping. Since the cloning of the molecular components of the CRAC channel, STIM1 and Orai1, FCDI of CRAC channel has been studied using HEK293T heterologous expression system. In this paper we describe a method of quantifying CRAC channel FCDI by using instantaneous tail currents.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Berridge MJ, Bootman MD, Roderick HL (2003) Calcium signalling: dynamics, homeostasis and remodelling. Nat Rev Mol Cell Biol 4:517–529

    Article  CAS  Google Scholar 

  2. Brehm P, Eckert R (1978) Calcium entry leads to inactivation of calcium channel in Paramecium. Science 202:1203–1206

    Article  CAS  PubMed  Google Scholar 

  3. Hering S, Berjukow S, Sokolov S et al (2000) Molecular determinants of inactivation in voltage-gated Ca2+ channels. J Physiol 528:237–249

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Fierro L, Parekh AB (1999) Fast calcium-dependent inactivation of calcium release-activated calcium current (CRAC) in RBL-1 cells. J Membr Biol 168:9–17

    Article  CAS  PubMed  Google Scholar 

  5. Hoth M, Penner R (1993) Calcium release-activated calcium current in rat mast cells. J Physiol 465:359–386

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Litjens T, Harland ML, Roberts ML et al (2004) Fast Ca2+-dependent inactivation of the store-operated Ca2+ current (ISOC) in liver cells: a role for calmodulin. J Physiol 558:85–97

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Zweifach A, Lewis RS (1995) Rapid inactivation of depletion-activated calcium current (ICRAC) due to local calcium feedback. J Gen Physiol 105:209–226

    Article  CAS  PubMed  Google Scholar 

  8. Scrimgeour NR, Wilson DP, Barritt GJ et al (2014) Structural and stoichiometric determinants of Ca2+ release-activated Ca2+ (CRAC) channel Ca2+-dependent inactivation. Biochim Biophys Acta 1838:1281–1287

    Article  CAS  PubMed  Google Scholar 

  9. Rychkov GY, Pusch M, Roberts ML et al (1998) Permeation and block of the skeletal muscle chloride channel, ClC-1, by foreign anions. J Gen Physiol 111:653–665

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Scrimgeour N, Litjens T, Ma L et al (2009) Properties of Orai1 mediated store-operated current depend on the expression levels of STIM1 and Orai1 proteins. J Physiol 587:2903–2918

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Hille B (2001) Ion channels of exitable membranes. Sinauer Associates, Sunderland, MA, pp 57–58

    Google Scholar 

  12. Hoover PJ, Lewis RS (2011) Stoichiometric requirements for trapping and gating of Ca2+ release-activated Ca2+ (CRAC) channels by stromal interaction molecule 1 (STIM1). Proc Natl Acad Sci U S A 108:13299–13304

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. School of Medicine, University of Adelaide, South Australian Health and Medical Research Institute (SAHMRI), Adelaide, SA, Australia

    Grigori Y. Rychkov

Authors
  1. Grigori Y. Rychkov
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Grigori Y. Rychkov .

Editor information

Editors and Affiliations

  1. IRSET Inserm UMR 1085/ UR1, Rennes, France

    Aubin Penna

  2. CNRS ERL 7003, Laboratoire STIM, Poitiers, France

    Bruno Constantin

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Rychkov, G.Y. (2018). Measurement of the CRAC Channel Fast Ca2+-Dependent Inactivation (FCDI). In: Penna, A., Constantin, B. (eds) The CRAC Channel. Methods in Molecular Biology, vol 1843. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8704-7_14

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-8704-7_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8702-3

  • Online ISBN: 978-1-4939-8704-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation