Visualisation and identification of the interaction between STIM1s in resting cells.

Store-operated Ca(2+) channels are a major Ca(2+) entry pathway in nonexcitable cells, which drive various essential cellular functions. Recently, STIM1 and Orai proteins have been identified as the major molecular components of the Ca(2+) release-activated Ca(2+) (CRAC) channel. As the key subunit...

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Autores principales:	Jun He, Tao Yu, Jingying Pan, He Li
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/1e47b31a088a4f86bb51c4ea280da2ca
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spelling	oai:doaj.org-article:1e47b31a088a4f86bb51c4ea280da2ca2021-11-18T07:25:00ZVisualisation and identification of the interaction between STIM1s in resting cells.1932-620310.1371/journal.pone.0033377https://doaj.org/article/1e47b31a088a4f86bb51c4ea280da2ca2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22438918/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Store-operated Ca(2+) channels are a major Ca(2+) entry pathway in nonexcitable cells, which drive various essential cellular functions. Recently, STIM1 and Orai proteins have been identified as the major molecular components of the Ca(2+) release-activated Ca(2+) (CRAC) channel. As the key subunit of the CRAC channel, STIM1 is the ER Ca(2+) sensor and is essential for the recruitment and activation of Orai1. However, the mechanisms in transmission of information of STIM1 to Orai1 still need further investigation. Bimolecular fluorescence complementation (BiFC) is one of the most advanced and powerful tools for studying and visualising protein-protein interactions in living cells. We utilised BiFC and acceptor photobleaching fluorescence resonance energy transfer (FRET) experiments to visualise and determine the state of STIM1 in the living cells in resting state. Our results demonstrate that STIM1 exists in an oligomeric form in resting cells and that rather than the SAM motif, it is the C-terminus (residues 233-474) of STIM1 that is the key domain for the interaction between STIM1s. The STIM1 oligomers (BiFC-STIM1) and wild-type STIM1 colocalised and had a fibrillar distribution in resting conditions. Depletion of ER Ca(2+) stores induced BiFC-STIM1 distribution to become punctate, an effect that could be prevented or reversed by 2-APB. After depletion of the Ca(2+) stores, BiFC-STIM1 has the ability to form puncta that colocalise with wild-type STIM1 or Orai1 near the plasma membrane. Our data also indicate that the function of BiFC-STIM1 was not altered compared with that of wild-type STIM1.Jun HeTao YuJingying PanHe LiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 3, p e33377 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jun He Tao Yu Jingying Pan He Li Visualisation and identification of the interaction between STIM1s in resting cells.
description	Store-operated Ca(2+) channels are a major Ca(2+) entry pathway in nonexcitable cells, which drive various essential cellular functions. Recently, STIM1 and Orai proteins have been identified as the major molecular components of the Ca(2+) release-activated Ca(2+) (CRAC) channel. As the key subunit of the CRAC channel, STIM1 is the ER Ca(2+) sensor and is essential for the recruitment and activation of Orai1. However, the mechanisms in transmission of information of STIM1 to Orai1 still need further investigation. Bimolecular fluorescence complementation (BiFC) is one of the most advanced and powerful tools for studying and visualising protein-protein interactions in living cells. We utilised BiFC and acceptor photobleaching fluorescence resonance energy transfer (FRET) experiments to visualise and determine the state of STIM1 in the living cells in resting state. Our results demonstrate that STIM1 exists in an oligomeric form in resting cells and that rather than the SAM motif, it is the C-terminus (residues 233-474) of STIM1 that is the key domain for the interaction between STIM1s. The STIM1 oligomers (BiFC-STIM1) and wild-type STIM1 colocalised and had a fibrillar distribution in resting conditions. Depletion of ER Ca(2+) stores induced BiFC-STIM1 distribution to become punctate, an effect that could be prevented or reversed by 2-APB. After depletion of the Ca(2+) stores, BiFC-STIM1 has the ability to form puncta that colocalise with wild-type STIM1 or Orai1 near the plasma membrane. Our data also indicate that the function of BiFC-STIM1 was not altered compared with that of wild-type STIM1.
format	article
author	Jun He Tao Yu Jingying Pan He Li
author_facet	Jun He Tao Yu Jingying Pan He Li
author_sort	Jun He
title	Visualisation and identification of the interaction between STIM1s in resting cells.
title_short	Visualisation and identification of the interaction between STIM1s in resting cells.
title_full	Visualisation and identification of the interaction between STIM1s in resting cells.
title_fullStr	Visualisation and identification of the interaction between STIM1s in resting cells.
title_full_unstemmed	Visualisation and identification of the interaction between STIM1s in resting cells.
title_sort	visualisation and identification of the interaction between stim1s in resting cells.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/1e47b31a088a4f86bb51c4ea280da2ca
work_keys_str_mv	AT junhe visualisationandidentificationoftheinteractionbetweenstim1sinrestingcells AT taoyu visualisationandidentificationoftheinteractionbetweenstim1sinrestingcells AT jingyingpan visualisationandidentificationoftheinteractionbetweenstim1sinrestingcells AT heli visualisationandidentificationoftheinteractionbetweenstim1sinrestingcells
_version_	1718423461081120768

Visualisation and identification of the interaction between STIM1s in resting cells.

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