A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity
TMEM16A forms calcium-activated chloride channels (CaCCs) that regulate physiological processes such as the secretions of airway epithelia and exocrine glands, the contraction of smooth muscles, and the excitability of neurons. Notwithstanding intense interest in the mechanism behind TMEM16A-CaCC ca...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2014-06-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/02772 |
| _version_ | 1811200333479673856 |
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| author | Jason Tien Christian J Peters Xiu Ming Wong Tong Cheng Yuh Nung Jan Lily Yeh Jan Huanghe Yang |
| author_facet | Jason Tien Christian J Peters Xiu Ming Wong Tong Cheng Yuh Nung Jan Lily Yeh Jan Huanghe Yang |
| author_sort | Jason Tien |
| collection | DOAJ |
| description | TMEM16A forms calcium-activated chloride channels (CaCCs) that regulate physiological processes such as the secretions of airway epithelia and exocrine glands, the contraction of smooth muscles, and the excitability of neurons. Notwithstanding intense interest in the mechanism behind TMEM16A-CaCC calcium-dependent gating, comprehensive surveys to identify and characterize potential calcium sensors of this channel are still lacking. By aligning distantly related calcium-activated ion channels in the TMEM16 family and conducting systematic mutagenesis of all conserved acidic residues thought to be exposed to the cytoplasm, we identify four acidic amino acids as putative calcium-binding residues. Alterations of the charge, polarity, and size of amino acid side chains at these sites alter the ability of different divalent cations to activate the channel. Furthermore, TMEM16A mutant channels containing double cysteine substitutions at these residues are sensitive to the redox potential of the internal solution, providing evidence for their physical proximity and solvent accessibility. |
| first_indexed | 2024-04-12T02:03:04Z |
| format | Article |
| id | doaj.art-fc977ecf315741d087276349126d515e |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T02:03:04Z |
| publishDate | 2014-06-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-fc977ecf315741d087276349126d515e2022-12-22T03:52:38ZengeLife Sciences Publications LtdeLife2050-084X2014-06-01310.7554/eLife.02772A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activityJason Tien0Christian J Peters1Xiu Ming Wong2Tong Cheng3Yuh Nung Jan4Lily Yeh Jan5Huanghe Yang6Department of Physiology, University of California, San Francisco, San Francisco, United StatesDepartment of Physiology, University of California, San Francisco, San Francisco, United StatesGraduate Program in Chemistry and Chemical Biology, University of California, San Francisco, San Francisco, United StatesDepartment of Physiology, University of California, San Francisco, San Francisco, United States; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States; Howard Hughes Medical Institute, University of California, San Fransisco, San Fransisco, United StatesDepartment of Physiology, University of California, San Francisco, San Francisco, United States; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States; Howard Hughes Medical Institute, University of California, San Fransisco, San Fransisco, United StatesDepartment of Physiology, University of California, San Francisco, San Francisco, United States; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States; Howard Hughes Medical Institute, University of California, San Fransisco, San Fransisco, United StatesDepartment of Physiology, University of California, San Francisco, San Francisco, United States; Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States; Howard Hughes Medical Institute, University of California, San Fransisco, San Fransisco, United StatesTMEM16A forms calcium-activated chloride channels (CaCCs) that regulate physiological processes such as the secretions of airway epithelia and exocrine glands, the contraction of smooth muscles, and the excitability of neurons. Notwithstanding intense interest in the mechanism behind TMEM16A-CaCC calcium-dependent gating, comprehensive surveys to identify and characterize potential calcium sensors of this channel are still lacking. By aligning distantly related calcium-activated ion channels in the TMEM16 family and conducting systematic mutagenesis of all conserved acidic residues thought to be exposed to the cytoplasm, we identify four acidic amino acids as putative calcium-binding residues. Alterations of the charge, polarity, and size of amino acid side chains at these sites alter the ability of different divalent cations to activate the channel. Furthermore, TMEM16A mutant channels containing double cysteine substitutions at these residues are sensitive to the redox potential of the internal solution, providing evidence for their physical proximity and solvent accessibility.https://elifesciences.org/articles/02772TMEM16 channelcalcium activated chloride channelcalcium binding siteTMEM16A channelANO1 channelcalcium dependent activation |
| spellingShingle | Jason Tien Christian J Peters Xiu Ming Wong Tong Cheng Yuh Nung Jan Lily Yeh Jan Huanghe Yang A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity eLife TMEM16 channel calcium activated chloride channel calcium binding site TMEM16A channel ANO1 channel calcium dependent activation |
| title | A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity |
| title_full | A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity |
| title_fullStr | A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity |
| title_full_unstemmed | A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity |
| title_short | A comprehensive search for calcium binding sites critical for TMEM16A calcium-activated chloride channel activity |
| title_sort | comprehensive search for calcium binding sites critical for tmem16a calcium activated chloride channel activity |
| topic | TMEM16 channel calcium activated chloride channel calcium binding site TMEM16A channel ANO1 channel calcium dependent activation |
| url | https://elifesciences.org/articles/02772 |
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