CRACM1 Multimers Form the Ion-Selective Pore of the CRAC Channel

Monika Vig, Andreas Beck, James M. Billingsley, Annette Lis, Suhel Parvez, Christine Peinelt, Dana L. Koomoa, Jonathan Soboloff, Donald Gill, Andrea Fleig, Jean Pierre Kinet, Reinhold Penner

Research output: Contribution to journalArticle

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Abstract

Receptor-mediated Ca2+ release from the endoplasmic reticulum (ER) is often followed by Ca2+ entry through Ca2+-release-activated Ca2+ (CRAC) channels in the plasma membrane [1-5]. RNAi screens have identified STIM1 as the putative ER Ca2+ sensor [6-8] and CRACM1 (Orai1; [9-11]) as the putative store-operated Ca2+ channel. Overexpression of both proteins is required to reconstitute CRAC currents (ICRAC; [11-14]). We show here that CRACM1 forms multimeric assemblies that bind STIM1 and that acidic residues in the transmembrane (TM) and extracellular domains of CRACM1 contribute to the ionic selectivity of the CRAC-channel pore. Replacement of the conserved glutamate in position 106 of the first TM domain of CRACM1 with glutamine (E106Q) acts as a dominant-negative protein, and substitution with aspartate (E106D) enhances Na+, Ba2+, and Sr2+ permeation relative to Ca2+. Mutating E190Q in TM3 also affects channel selectivity, suggesting that glutamate residues in both TM1 and TM3 face the lumen of the pore. Furthermore, mutating a putative Ca2+ binding site in the first extracellular loop of CRACM1 (D110/112A) enhances monovalent cation permeation, suggesting that these residues too contribute to the coordination of Ca2+ ions to the pore. Our data provide unequivocal evidence that CRACM1 multimers form the Ca2+-selective CRAC-channel pore.

Original languageEnglish (US)
Pages (from-to)2073-2079
Number of pages7
JournalCurrent Biology
Volume16
Issue number20
DOIs
StatePublished - Oct 24 2006

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Permeation
Endoplasmic Reticulum
Glutamic Acid
Ions
ions
calcium
Monovalent Cations
Cell membranes
RNA Interference
Glutamine
Aspartic Acid
Proteins
Substitution reactions
Binding Sites
Cell Membrane
Sensors
glutamates
endoplasmic reticulum
aspartic acid
glutamine

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Vig, M., Beck, A., Billingsley, J. M., Lis, A., Parvez, S., Peinelt, C., ... Penner, R. (2006). CRACM1 Multimers Form the Ion-Selective Pore of the CRAC Channel. Current Biology, 16(20), 2073-2079. https://doi.org/10.1016/j.cub.2006.08.085
Vig, Monika ; Beck, Andreas ; Billingsley, James M. ; Lis, Annette ; Parvez, Suhel ; Peinelt, Christine ; Koomoa, Dana L. ; Soboloff, Jonathan ; Gill, Donald ; Fleig, Andrea ; Kinet, Jean Pierre ; Penner, Reinhold. / CRACM1 Multimers Form the Ion-Selective Pore of the CRAC Channel. In: Current Biology. 2006 ; Vol. 16, No. 20. pp. 2073-2079.
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Vig, M, Beck, A, Billingsley, JM, Lis, A, Parvez, S, Peinelt, C, Koomoa, DL, Soboloff, J, Gill, D, Fleig, A, Kinet, JP & Penner, R 2006, 'CRACM1 Multimers Form the Ion-Selective Pore of the CRAC Channel', Current Biology, vol. 16, no. 20, pp. 2073-2079. https://doi.org/10.1016/j.cub.2006.08.085

CRACM1 Multimers Form the Ion-Selective Pore of the CRAC Channel. / Vig, Monika; Beck, Andreas; Billingsley, James M.; Lis, Annette; Parvez, Suhel; Peinelt, Christine; Koomoa, Dana L.; Soboloff, Jonathan; Gill, Donald; Fleig, Andrea; Kinet, Jean Pierre; Penner, Reinhold.

In: Current Biology, Vol. 16, No. 20, 24.10.2006, p. 2073-2079.

Research output: Contribution to journalArticle

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T1 - CRACM1 Multimers Form the Ion-Selective Pore of the CRAC Channel

AU - Vig, Monika

AU - Beck, Andreas

AU - Billingsley, James M.

AU - Lis, Annette

AU - Parvez, Suhel

AU - Peinelt, Christine

AU - Koomoa, Dana L.

AU - Soboloff, Jonathan

AU - Gill, Donald

AU - Fleig, Andrea

AU - Kinet, Jean Pierre

AU - Penner, Reinhold

PY - 2006/10/24

Y1 - 2006/10/24

N2 - Receptor-mediated Ca2+ release from the endoplasmic reticulum (ER) is often followed by Ca2+ entry through Ca2+-release-activated Ca2+ (CRAC) channels in the plasma membrane [1-5]. RNAi screens have identified STIM1 as the putative ER Ca2+ sensor [6-8] and CRACM1 (Orai1; [9-11]) as the putative store-operated Ca2+ channel. Overexpression of both proteins is required to reconstitute CRAC currents (ICRAC; [11-14]). We show here that CRACM1 forms multimeric assemblies that bind STIM1 and that acidic residues in the transmembrane (TM) and extracellular domains of CRACM1 contribute to the ionic selectivity of the CRAC-channel pore. Replacement of the conserved glutamate in position 106 of the first TM domain of CRACM1 with glutamine (E106Q) acts as a dominant-negative protein, and substitution with aspartate (E106D) enhances Na+, Ba2+, and Sr2+ permeation relative to Ca2+. Mutating E190Q in TM3 also affects channel selectivity, suggesting that glutamate residues in both TM1 and TM3 face the lumen of the pore. Furthermore, mutating a putative Ca2+ binding site in the first extracellular loop of CRACM1 (D110/112A) enhances monovalent cation permeation, suggesting that these residues too contribute to the coordination of Ca2+ ions to the pore. Our data provide unequivocal evidence that CRACM1 multimers form the Ca2+-selective CRAC-channel pore.

AB - Receptor-mediated Ca2+ release from the endoplasmic reticulum (ER) is often followed by Ca2+ entry through Ca2+-release-activated Ca2+ (CRAC) channels in the plasma membrane [1-5]. RNAi screens have identified STIM1 as the putative ER Ca2+ sensor [6-8] and CRACM1 (Orai1; [9-11]) as the putative store-operated Ca2+ channel. Overexpression of both proteins is required to reconstitute CRAC currents (ICRAC; [11-14]). We show here that CRACM1 forms multimeric assemblies that bind STIM1 and that acidic residues in the transmembrane (TM) and extracellular domains of CRACM1 contribute to the ionic selectivity of the CRAC-channel pore. Replacement of the conserved glutamate in position 106 of the first TM domain of CRACM1 with glutamine (E106Q) acts as a dominant-negative protein, and substitution with aspartate (E106D) enhances Na+, Ba2+, and Sr2+ permeation relative to Ca2+. Mutating E190Q in TM3 also affects channel selectivity, suggesting that glutamate residues in both TM1 and TM3 face the lumen of the pore. Furthermore, mutating a putative Ca2+ binding site in the first extracellular loop of CRACM1 (D110/112A) enhances monovalent cation permeation, suggesting that these residues too contribute to the coordination of Ca2+ ions to the pore. Our data provide unequivocal evidence that CRACM1 multimers form the Ca2+-selective CRAC-channel pore.

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Vig M, Beck A, Billingsley JM, Lis A, Parvez S, Peinelt C et al. CRACM1 Multimers Form the Ion-Selective Pore of the CRAC Channel. Current Biology. 2006 Oct 24;16(20):2073-2079. https://doi.org/10.1016/j.cub.2006.08.085