Expression and preliminary characterization of human MICU2
MICU2 has been reported to interact with MICU1 and participate in the regulation of mitochondrial Ca2+ uptake, although the molecular determinants underlying the function of MICU2 is unknown. In order to characterize MICU2 we screened a series of N-terminal and C-terminal truncations and obtained co...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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The Company of Biologists
2016-07-01
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| Series: | Biology Open |
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| Online Access: | http://bio.biologists.org/content/5/7/962 |
| _version_ | 1818859663196684288 |
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| author | Dan Li Wenping Wu Hairun Pei Qiang Wei Qingzhan Yang Jimin Zheng Zongchao Jia |
| author_facet | Dan Li Wenping Wu Hairun Pei Qiang Wei Qingzhan Yang Jimin Zheng Zongchao Jia |
| author_sort | Dan Li |
| collection | DOAJ |
| description | MICU2 has been reported to interact with MICU1 and participate in the regulation of mitochondrial Ca2+ uptake, although the molecular determinants underlying the function of MICU2 is unknown. In order to characterize MICU2 we screened a series of N-terminal and C-terminal truncations and obtained constructs which can be expressed in abundance, giving rise to soluble samples to enable subsequent characterizations. Size exclusion chromatography (SEC) and multi-angle laser light scattering (MALLS) revealed that MICU2 exists as a monomer in Ca2+-free conditions but forms a dimer in Ca2+-bound conditions. Unlike MICU1, the C-helix domain of MICU2 exhibits no influence on protein conformation in both Ca2+-free and Ca2+-bound forms. Furthermore, mutation of the first EF-hand abolishes the ability of MICU2 to switch to a dimer in the presence of Ca2+, indicating that the first EF-hand is not only involved in Ca2+ binding but also in conformational change. Our pull-down and co-immunoprecipitation assays suggest that, in addition to disulfide bonds, salt bridges also contribute to MICU1-MICU2 heterodimer formation. |
| first_indexed | 2024-12-19T09:15:45Z |
| format | Article |
| id | doaj.art-27aaa4a21d71426a95b4218d6f3d6cb4 |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-19T09:15:45Z |
| publishDate | 2016-07-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-27aaa4a21d71426a95b4218d6f3d6cb42022-12-21T20:28:05ZengThe Company of BiologistsBiology Open2046-63902016-07-015796296910.1242/bio.018572018572Expression and preliminary characterization of human MICU2Dan Li0Wenping Wu1Hairun Pei2Qiang Wei3Qingzhan Yang4Jimin Zheng5Zongchao Jia6 College of Chemistry, Beijing Normal University, Beijing 100875, China College of Chemistry, Beijing Normal University, Beijing 100875, China College of Chemistry, Beijing Normal University, Beijing 100875, China College of Chemistry, Beijing Normal University, Beijing 100875, China College of Chemistry, Beijing Normal University, Beijing 100875, China College of Chemistry, Beijing Normal University, Beijing 100875, China Department of Biochemical and Molecular Science, Queen University, Kingston, Ontario, Canada K7L 3N6 MICU2 has been reported to interact with MICU1 and participate in the regulation of mitochondrial Ca2+ uptake, although the molecular determinants underlying the function of MICU2 is unknown. In order to characterize MICU2 we screened a series of N-terminal and C-terminal truncations and obtained constructs which can be expressed in abundance, giving rise to soluble samples to enable subsequent characterizations. Size exclusion chromatography (SEC) and multi-angle laser light scattering (MALLS) revealed that MICU2 exists as a monomer in Ca2+-free conditions but forms a dimer in Ca2+-bound conditions. Unlike MICU1, the C-helix domain of MICU2 exhibits no influence on protein conformation in both Ca2+-free and Ca2+-bound forms. Furthermore, mutation of the first EF-hand abolishes the ability of MICU2 to switch to a dimer in the presence of Ca2+, indicating that the first EF-hand is not only involved in Ca2+ binding but also in conformational change. Our pull-down and co-immunoprecipitation assays suggest that, in addition to disulfide bonds, salt bridges also contribute to MICU1-MICU2 heterodimer formation.http://bio.biologists.org/content/5/7/962MICU1MICU2CalciumEF-handMitochondria |
| spellingShingle | Dan Li Wenping Wu Hairun Pei Qiang Wei Qingzhan Yang Jimin Zheng Zongchao Jia Expression and preliminary characterization of human MICU2 Biology Open MICU1 MICU2 Calcium EF-hand Mitochondria |
| title | Expression and preliminary characterization of human MICU2 |
| title_full | Expression and preliminary characterization of human MICU2 |
| title_fullStr | Expression and preliminary characterization of human MICU2 |
| title_full_unstemmed | Expression and preliminary characterization of human MICU2 |
| title_short | Expression and preliminary characterization of human MICU2 |
| title_sort | expression and preliminary characterization of human micu2 |
| topic | MICU1 MICU2 Calcium EF-hand Mitochondria |
| url | http://bio.biologists.org/content/5/7/962 |
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