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Molecular mechanisms underlying inhibition of STIM1-Orai1-mediated Ca2+ entry induced by 2-aminoethoxydiphenyl borate.
Pflugers Arch 2016; 468(11-12):2061-2074PA

Abstract

Store-operated Ca2+ entry (SOCE) mediated by STIM1 and Orai1 is crucial for Ca2+ signaling and homeostasis in most cell types. 2-Aminoethoxydiphenyl borate (2-APB) is a well-described SOCE inhibitor, but its mechanisms of action remain largely elusive. Here, we show that 2-APB does not affect the dimeric state of STIM1, but enhances the intramolecular coupling between the coiled-coil 1 (CC1) and STIM-Orai-activating region (SOAR) of STIM1, with subsequent reduction in the formation of STIM1 puncta in the absence of Orai1 overexpression. 2-APB also inhibits Orai1 channels, directly inhibiting Ca2+ entry through the constitutively active, STIM1-independent Orai1 mutants, Orai1-P245T and Orai1-V102A. When unbound from STIM1, the constitutively active Orai1-V102C mutant is not inhibited by 2-APB. Thus, we used Orai1-V012C as a tool to examine whether 2-APB can also inhibit the coupling between STIM1 and Orai1. We reveal that the functional coupling between STIM1 and Orai1-V102C is inhibited by 2-APB. This inhibition on coupling is indirect, arising from 2-APB's action on STIM1, and it is most likely mediated by functional channel residues in the Orai1 N-terminus. Overall, our findings on this two-site inhibition mediated by 2-APB provide new understanding on Orai1-activation by STIM1, important to future drug design.

Authors+Show Affiliations

Beijing Key Laboratory of Gene Resources and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, People's Republic of China.Department of Cellular and Molecular Physiology, The Pennsylvania State University College of Medicine, Hershey, PA, 17033, USA.Beijing Key Laboratory of Gene Resources and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, People's Republic of China.Institute of Biosciences and Technology, Texas A&M University Health Science Center, Houston, TX, 77030, USA.Institute of Biosciences and Technology, Texas A&M University Health Science Center, Houston, TX, 77030, USA.Beijing Key Laboratory of Gene Resources and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, People's Republic of China.Beijing Key Laboratory of Gene Resources and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, People's Republic of China.Beijing Key Laboratory of Gene Resources and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, People's Republic of China.Department of Medical Physiology, College of Medicine, Texas A&M University Health Science Center, Temple, TX, 76504, USA.Department of Cellular and Molecular Physiology, The Pennsylvania State University College of Medicine, Hershey, PA, 17033, USA. dgill4@hmc.psu.edu.Beijing Key Laboratory of Gene Resources and Molecular Development, College of Life Sciences, Beijing Normal University, Beijing, 100875, People's Republic of China. wyoujun@bnu.edu.cn.

Pub Type(s)

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

Language

eng

PubMed ID

27726010

Citation

Wei, Ming, et al. "Molecular Mechanisms Underlying Inhibition of STIM1-Orai1-mediated Ca2+ Entry Induced By 2-aminoethoxydiphenyl Borate." Pflugers Archiv : European Journal of Physiology, vol. 468, no. 11-12, 2016, pp. 2061-2074.
Wei M, Zhou Y, Sun A, et al. Molecular mechanisms underlying inhibition of STIM1-Orai1-mediated Ca2+ entry induced by 2-aminoethoxydiphenyl borate. Pflugers Arch. 2016;468(11-12):2061-2074.
Wei, M., Zhou, Y., Sun, A., Ma, G., He, L., Zhou, L., ... Wang, Y. (2016). Molecular mechanisms underlying inhibition of STIM1-Orai1-mediated Ca2+ entry induced by 2-aminoethoxydiphenyl borate. Pflugers Archiv : European Journal of Physiology, 468(11-12), pp. 2061-2074.
Wei M, et al. Molecular Mechanisms Underlying Inhibition of STIM1-Orai1-mediated Ca2+ Entry Induced By 2-aminoethoxydiphenyl Borate. Pflugers Arch. 2016;468(11-12):2061-2074. PubMed PMID: 27726010.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular mechanisms underlying inhibition of STIM1-Orai1-mediated Ca2+ entry induced by 2-aminoethoxydiphenyl borate. AU - Wei,Ming, AU - Zhou,Yandong, AU - Sun,Aomin, AU - Ma,Guolin, AU - He,Lian, AU - Zhou,Lijuan, AU - Zhang,Shuce, AU - Liu,Jin, AU - Zhang,Shenyuan L, AU - Gill,Donald L, AU - Wang,Youjun, Y1 - 2016/10/10/ PY - 2016/06/22/received PY - 2016/09/07/accepted PY - 2016/08/24/revised PY - 2016/10/12/pubmed PY - 2017/7/1/medline PY - 2016/10/12/entrez KW - 2-APB KW - Calcium KW - FRET KW - Orai1 KW - Puncta KW - SOCE KW - STIM1 SP - 2061 EP - 2074 JF - Pflugers Archiv : European journal of physiology JO - Pflugers Arch. VL - 468 IS - 11-12 N2 - Store-operated Ca2+ entry (SOCE) mediated by STIM1 and Orai1 is crucial for Ca2+ signaling and homeostasis in most cell types. 2-Aminoethoxydiphenyl borate (2-APB) is a well-described SOCE inhibitor, but its mechanisms of action remain largely elusive. Here, we show that 2-APB does not affect the dimeric state of STIM1, but enhances the intramolecular coupling between the coiled-coil 1 (CC1) and STIM-Orai-activating region (SOAR) of STIM1, with subsequent reduction in the formation of STIM1 puncta in the absence of Orai1 overexpression. 2-APB also inhibits Orai1 channels, directly inhibiting Ca2+ entry through the constitutively active, STIM1-independent Orai1 mutants, Orai1-P245T and Orai1-V102A. When unbound from STIM1, the constitutively active Orai1-V102C mutant is not inhibited by 2-APB. Thus, we used Orai1-V012C as a tool to examine whether 2-APB can also inhibit the coupling between STIM1 and Orai1. We reveal that the functional coupling between STIM1 and Orai1-V102C is inhibited by 2-APB. This inhibition on coupling is indirect, arising from 2-APB's action on STIM1, and it is most likely mediated by functional channel residues in the Orai1 N-terminus. Overall, our findings on this two-site inhibition mediated by 2-APB provide new understanding on Orai1-activation by STIM1, important to future drug design. SN - 1432-2013 UR - https://www.unboundmedicine.com/medline/citation/27726010/Molecular_mechanisms_underlying_inhibition_of_STIM1_Orai1_mediated_Ca2+_entry_induced_by_2_aminoethoxydiphenyl_borate_ L2 - https://dx.doi.org/10.1007/s00424-016-1880-z DB - PRIME DP - Unbound Medicine ER -