Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.


Logo for

J. Biol. Chem. 282 (40): 29678-29690

© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.

Visualization and Manipulation of Plasma Membrane-Endoplasmic Reticulum Contact Sites Indicates the Presence of Additional Molecular Components within the STIM1-Orai1 Complex*



Péter Várnai{ddagger}§1, Balázs Tóth{ddagger}, Dániel J. Tóth§, László Hunyady§, , and Tamas Balla{ddagger}2

{ddagger}Section on Molecular Signal Transduction, NICHD, National Institutes of Health, Bethesda, Maryland 20892-4510 and the §Department of Physiology, Semmelweis University, School of Medicine, Budapest, Hungary, H-1086

Abstract: STIM1, a recently identified endoplasmic reticulum (ER) protein, rapidly translocates to a plasma membrane-adjacent ER compartment upon depletion of the ER Ca2+ stores. Here we use a novel means, namely a chemically inducible bridge formation between the plasma and ER membranes, to highlight the plasma membrane-adjacent ER compartment and show that this is the site where STIM1 and its Ca2+ channel partner, Orai1, form a productive interaction upon store depletion. By changing the length of the linkers connecting the plasma and ER membranes, we show that Orai1 requires a larger space than STIM1 between the two membranes. This finding suggests that Orai1 is part of a larger macromolecular cluster with an estimated 11-14-nm protrusion to the cytoplasm, whereas the cytoplasmic domain of STIM1 fits in a space calculated to be less than 6 nm. We finally show that agonist-induced translocation of STIM1 is rapidly reversible and only partially affects STIM1 in the juxtanuclear ER compartment. These studies are the first to detect juxtaposed areas between the ER and the plasma membrane in live cells, revealing novel details of STIM1-Orai1 interactions.

Received for publication May 25, 2007. Revision received July 6, 2007.

* This work was supported in part by the Intramural Research Program of the National Institute of Child Health and Human Development of the National Institutes of Health (to B. T., P. V., and T. B.) and by an appointment of PV to the Senior Fellowship Program at the NIH. This latter program is administered by the Oak Ridge Institute for Science and Education through an interagency agreement between the U.S. Department of Energy and the National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.


The on-line version of this article (available at contains three supplemental figures and three movies.

{diamondsuit} This article was selected as a Paper of the Week.

1 A Bolyai Fellow of the Hungarian Academy of Science. Supported by the Hungarian Scientific Research fund (Grant OTKA NF-68563) and the Medical Research Council (Grant ETT 440/2006).

2 To whom correspondence should be addressed: National Institutes of Health, Bldg. 49, Rm. 6A35, 49 Convent Dr., Bethesda, MD 20892-4510. Tel.: 301-496-2136; Fax: 301-480-8010; E-mail: ballat{at}

STARD3 or STARD3NL and VAP form a novel molecular tether between late endosomes and the ER.
F. Alpy, A. Rousseau, Y. Schwab, F. Legueux, I. Stoll, C. Wendling, C. Spiegelhalter, P. Kessler, C. Mathelin, M.-C. Rio, et al. (2013)
J. Cell Sci. 126, 5500-5512
   Abstract »    Full Text »    PDF »
Phosphoinositides: Tiny Lipids With Giant Impact on Cell Regulation.
T. Balla (2013)
Physiol Rev 93, 1019-1137
   Abstract »    Full Text »    PDF »
Role of Phospholipids in Endocytosis, Phagocytosis, and Macropinocytosis.
M. Bohdanowicz and S. Grinstein (2013)
Physiol Rev 93, 69-106
   Abstract »    Full Text »    PDF »
An essential and NSF independent role for {alpha}-SNAP in store-operated calcium entry.
Y. Miao, C. Miner, L. Zhang, P. I. Hanson, A. Dani, and M. Vig (2013)
eLife Sci 2, e00802
   Abstract »    Full Text »    PDF »
{sigma}-1 Receptor at the Mitochondrial-Associated Endoplasmic Reticulum Membrane Is Responsible for Mitochondrial Metabolic Regulation.
K.-S. C. Marriott, M. Prasad, V. Thapliyal, and H. S. Bose (2012)
J. Pharmacol. Exp. Ther. 343, 578-586
   Abstract »    Full Text »    PDF »
Bidirectional Coupling between Ryanodine Receptors and Ca2+ Release-activated Ca2+ (CRAC) Channel Machinery Sustains Store-operated Ca2+ Entry in Human T Lymphocytes.
P. Thakur, S. Dadsetan, and A. F. Fomina (2012)
J. Biol. Chem. 287, 37233-37244
   Abstract »    Full Text »    PDF »
Orai1, STIM1, and their associating partners.
S. Srikanth and Y. Gwack (2012)
J. Physiol. 590, 4169-4177
   Abstract »    Full Text »    PDF »
Overexpression of Orai1 and STIM1 Proteins Alters Regulation of Store-operated Ca2+ Entry by Endogenous Mediators.
T. Gwozdz, J. Dutko-Gwozdz, C. Schafer, and V. M. Bolotina (2012)
J. Biol. Chem. 287, 22865-22872
   Abstract »    Full Text »    PDF »
Junctate is a Ca2+-sensing structural component of Orai1 and stromal interaction molecule 1 (STIM1).
S. Srikanth, M. Jew, K.-D. Kim, M.-K. Yee, J. Abramson, and Y. Gwack (2012)
PNAS 109, 8682-8687
   Abstract »    Full Text »    PDF »
Acute depletion of plasma membrane phosphatidylinositol 4,5-bisphosphate impairs specific steps in endocytosis of the G-protein-coupled receptor.
D. J. Toth, J. T. Toth, G. Gulyas, A. Balla, T. Balla, L. Hunyady, and P. Varnai (2012)
J. Cell Sci. 125, 2185-2197
   Abstract »    Full Text »    PDF »
Urotensin-II Signaling Mechanism in Rat Coronary Artery: Role of STIM1 and Orai1-Dependent Store Operated Calcium Influx in Vasoconstriction.
A. Dominguez-Rodriguez, I. Diaz, M. Rodriguez-Moyano, E. Calderon-Sanchez, J. A. Rosado, A. Ordonez, and T. Smani (2012)
Arterioscler Thromb Vasc Biol 32, 1325-1332
   Abstract »    Full Text »    PDF »
Perispeckles are major assembly sites for the exon junction core complex.
E. Daguenet, A. Baguet, S. Degot, U. Schmidt, F. Alpy, C. Wendling, C. Spiegelhalter, P. Kessler, M.-C. Rio, H. Le Hir, et al. (2012)
Mol. Biol. Cell 23, 1765-1782
   Abstract »    Full Text »    PDF »
Two phosphatidylinositol 4-kinases control lysosomal delivery of the Gaucher disease enzyme, {beta}-glucocerebrosidase.
M. Jovic, M. J. Kean, Z. Szentpetery, G. Polevoy, A.-C. Gingras, J. A. Brill, and T. Balla (2012)
Mol. Biol. Cell 23, 1533-1545
   Abstract »    Full Text »    PDF »
Molecular Mechanism of Pancreatic and Salivary Gland Fluid and HCOFormula Secretion.
M. G. Lee, E. Ohana, H. W. Park, D. Yang, and S. Muallem (2012)
Physiol Rev 92, 39-74
   Abstract »    Full Text »    PDF »
Store-Operated Calcium Channels: New Perspectives on Mechanism and Function.
R. S. Lewis (2011)
Cold Spring Harb Perspect Biol 3, a003970
   Abstract »    Full Text »    PDF »
A single lysine in the N-terminal region of store-operated channels is critical for STIM1-mediated gating.
A. Lis, S. Zierler, C. Peinelt, A. Fleig, and R. Penner (2010)
J. Gen. Physiol. 136, 673-686
   Abstract »    Full Text »    PDF »
Agonist-activated Ca2+ influx occurs at stable plasma membrane and endoplasmic reticulum junctions.
S. Treves, M. Vukcevic, J. Griesser, C.-F. Armstrong, M. X. Zhu, and F. Zorzato (2010)
J. Cell Sci. 123, 4170-4181
   Abstract »    Full Text »    PDF »
Activation of STIM1-Orai1 Involves an Intramolecular Switching Mechanism.
M. K. Korzeniowski, I. M. Manjarres, P. Varnai, and T. Balla (2010)
Science Signaling 3, ra82
   Abstract »    Full Text »    PDF »
Orai1 interacts with STIM1 and mediates capacitative Ca2+ entry in mouse pulmonary arterial smooth muscle cells.
L. C. Ng, D. Ramduny, J. A. Airey, C. A. Singer, P. S. Keller, X.-M. Shen, H. Tian, M. Valencik, and J. R. Hume (2010)
Am J Physiol Cell Physiol 299, C1079-C1090
   Abstract »    Full Text »    PDF »
Acetylsalicylic acid enhances purinergic receptor-mediated outward currents in rat megakaryocytes.
J. P. Young, J. Beckerman, S. Vicini, and A. Myers (2010)
Am J Physiol Cell Physiol 298, C602-C610
   Abstract »    Full Text »    PDF »
Lipid-regulated sterol transfer between closely apposed membranes by oxysterol-binding protein homologues.
T. A. Schulz, M.-G. Choi, S. Raychaudhuri, J. A. Mears, R. Ghirlando, J. E. Hinshaw, and W. A. Prinz (2009)
J. Cell Biol. 187, 889-903
   Abstract »    Full Text »    PDF »
STIM1-induced precortical and cortical subdomains of the endoplasmic reticulum.
L. Orci, M. Ravazzola, M. Le Coadic, W.-w. Shen, N. Demaurex, and P. Cosson (2009)
PNAS 106, 19358-19362
   Abstract »    Full Text »    PDF »
STIM and Orai: Dynamic Intermembrane Coupling to Control Cellular Calcium Signals.
X. Deng, Y. Wang, Y. Zhou, J. Soboloff, and D. L. Gill (2009)
J. Biol. Chem. 284, 22501-22505
   Abstract »    Full Text »    PDF »
Molecular Determinants of the Coupling between STIM1 and Orai Channels: DIFFERENTIAL ACTIVATION OF Orai1-3 CHANNELS BY A STIM1 COILED-COIL MUTANT.
I. Frischauf, M. Muik, I. Derler, J. Bergsmann, M. Fahrner, R. Schindl, K. Groschner, and C. Romanin (2009)
J. Biol. Chem. 284, 21696-21706
   Abstract »    Full Text »    PDF »
Phosphoinositide Signaling: New Tools and Insights.
T. Balla, Z. Szentpetery, and Y. J. Kim (2009)
Physiology 24, 231-244
   Abstract »    Full Text »    PDF »
Dependence of STIM1/Orai1-mediated Calcium Entry on Plasma Membrane Phosphoinositides.
M. K. Korzeniowski, M. A. Popovic, Z. Szentpetery, P. Varnai, S. S. Stojilkovic, and T. Balla (2009)
J. Biol. Chem. 284, 21027-21035
   Abstract »    Full Text »    PDF »
Properties of Orai1 mediated store-operated current depend on the expression levels of STIM1 and Orai1 proteins.
N. Scrimgeour, T. Litjens, L. Ma, G. J. Barritt, and G. Y. Rychkov (2009)
J. Physiol. 587, 2903-2918
   Abstract »    Full Text »    PDF »
Tetrameric Orai1 Is a Teardrop-shaped Molecule with a Long, Tapered Cytoplasmic Domain.
Y. Maruyama, T. Ogura, K. Mio, K. Kato, T. Kaneko, S. Kiyonaka, Y. Mori, and C. Sato (2009)
J. Biol. Chem. 284, 13676-13685
   Abstract »    Full Text »    PDF »
Molecular Clustering of STIM1 with Orai1/CRACM1 at the Plasma Membrane Depends Dynamically on Depletion of Ca2+ Stores and on Electrostatic Interactions.
N. Calloway, M. Vig, J.-P. Kinet, D. Holowka, and B. Baird (2009)
Mol. Biol. Cell 20, 389-399
   Abstract »    Full Text »    PDF »
How strict is the correlation between STIM1 and Orai1 expression, puncta formation, and ICRAC activation?.
T. Gwozdz, J. Dutko-Gwozdz, V. Zarayskiy, K. Peter, and V. M. Bolotina (2008)
Am J Physiol Cell Physiol 295, C1133-C1140
   Abstract »    Full Text »    PDF »
Selective coupling of type 6 adenylyl cyclase with type 2 IP3 receptors mediates direct sensitization of IP3 receptors by cAMP.
S. C. Tovey, S. G. Dedos, E. J.A. Taylor, J. E. Church, and C. W. Taylor (2008)
J. Cell Biol. 183, 297-311
   Abstract »    Full Text »    PDF »
Location and Function of STIM1 in the Activation of Ca2+ Entry Signals.
T. Hewavitharana, X. Deng, Y. Wang, M. F. Ritchie, G. V. Girish, J. Soboloff, and D. L. Gill (2008)
J. Biol. Chem. 283, 26252-26262
   Abstract »    Full Text »    PDF »
Functional stoichiometry of the unitary calcium-release-activated calcium channel.
W. Ji, P. Xu, Z. Li, J. Lu, L. Liu, Y. Zhan, Y. Chen, B. Hille, T. Xu, and L. Chen (2008)
PNAS 105, 13668-13673
   Abstract »    Full Text »    PDF »
Orai, STIM1 and iPLA2{beta}: a view from a different perspective.
V. M. Bolotina (2008)
J. Physiol. 586, 3035-3042
   Abstract »    Full Text »    PDF »
Dynamic Movement of the Calcium Sensor STIM1 and the Calcium Channel Orai1 in Activated T-Cells: Puncta and Distal Caps.
V. A. Barr, K. M. Bernot, S. Srikanth, Y. Gwack, L. Balagopalan, C. K. Regan, D. J. Helman, C. L. Sommers, M. Oh-hora, A. Rao, et al. (2008)
Mol. Biol. Cell 19, 2802-2817
   Abstract »    Full Text »    PDF »
Lipid Rafts Determine Clustering of STIM1 in Endoplasmic Reticulum-Plasma Membrane Junctions and Regulation of Store-operated Ca2+ Entry (SOCE).
B. Pani, H. L. Ong, X. Liu, K. Rauser, I. S. Ambudkar, and B. B. Singh (2008)
J. Biol. Chem. 283, 17333-17340
   Abstract »    Full Text »    PDF »
Voltage Gating at the Selectivity Filter of the Ca2+ Release-activated Ca2+ Channel Induced by Mutation of the Orai1 Protein.
M. A. Spassova, T. Hewavitharana, R. A. Fandino, A. Kaya, J. Tanaka, and D. L. Gill (2008)
J. Biol. Chem. 283, 14938-14945
   Abstract »    Full Text »    PDF »
Novel Role for STIM1 as a Trigger for Calcium Influx Factor Production.
P. Csutora, K. Peter, H. Kilic, K. M. Park, V. Zarayskiy, T. Gwozdz, and V. M. Bolotina (2008)
J. Biol. Chem. 283, 14524-14531
   Abstract »    Full Text »    PDF »
Functional Requirement for Orai1 in Store-operated TRPC1-STIM1 Channels.
K. T. Cheng, X. Liu, H. L. Ong, and I. S. Ambudkar (2008)
J. Biol. Chem. 283, 12935-12940
   Abstract »    Full Text »    PDF »
Dynamic Coupling of the Putative Coiled-coil Domain of ORAI1 with STIM1 Mediates ORAI1 Channel Activation.
M. Muik, I. Frischauf, I. Derler, M. Fahrner, J. Bergsmann, P. Eder, R. Schindl, C. Hesch, B. Polzinger, R. Fritsch, et al. (2008)
J. Biol. Chem. 283, 8014-8022
   Abstract »    Full Text »    PDF »
Ca2+-store-dependent and -independent reversal of Stim1 localization and function.
J. T. Smyth, W. I. DeHaven, G. S. Bird, and J. W. Putney Jr (2008)
J. Cell Sci. 121, 762-772
   Abstract »    Full Text »    PDF »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882