Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells
Small heat shock proteins (sHsps) endow cells with stress tolerance. Of the various sHsps in mammals, HspB1, also known as Hsp27, is the most ubiquitous. To examine the structure and function of HspB1, we expressed, purified, and characterized HspB1 from Chinese hamster (Cricetulus griseus) ovary ce...
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| Format: | Article |
| Language: | English |
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Wiley
2019-10-01
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| Series: | FEBS Open Bio |
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| Online Access: | https://doi.org/10.1002/2211-5463.12726 |
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| author | Eiryo Sha Manami Nakamura Kazuya Ankai Yohei Y. Yamamoto Toshihiko Oka Masafumi Yohda |
| author_facet | Eiryo Sha Manami Nakamura Kazuya Ankai Yohei Y. Yamamoto Toshihiko Oka Masafumi Yohda |
| author_sort | Eiryo Sha |
| collection | DOAJ |
| description | Small heat shock proteins (sHsps) endow cells with stress tolerance. Of the various sHsps in mammals, HspB1, also known as Hsp27, is the most ubiquitous. To examine the structure and function of HspB1, we expressed, purified, and characterized HspB1 from Chinese hamster (Cricetulus griseus) ovary cells (CgHspB1). CgHspB1 forms a large oligomeric structure. We observed a monodisperse 16‐mer with an elongated sphere, but this is affected by changes in various conditions, including temperature. Under dilute conditions, CgHspB1 dissociates into small oligomers at elevated temperatures. The dissociated conformers interacted with the gel filtration column through hydrophobic interactions. In contrast, dissociation of the oligomer was not observed by small‐angle X‐ray scattering at 55 °C. The result partially coincides with the results of size exclusion chromatography, showing that dissociation did not occur at high protein concentrations. However, a significant structural change in the oligomeric conformations appears to occur between room and higher temperatures. Reflecting their status as homeotherms, mammalian sHsps are regulated by phosphorylation. A phosphorylation mimic mutant of CgHspB1 with the replacement of Ser15 to Asp exhibited relatively lower oligomer stability and greater protective ability against thermal aggregation than the wild‐type protein. The result clearly shows a correlation between oligomer dissociation and chaperone activity. |
| format | Article |
| id | doaj-art-aeadb2a6784c49e091b998437b098e00 |
| institution | Kabale University |
| issn | 2211-5463 |
| language | English |
| publishDate | 2019-10-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj-art-aeadb2a6784c49e091b998437b098e002024-12-06T11:26:04ZengWileyFEBS Open Bio2211-54632019-10-019101826183410.1002/2211-5463.12726Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cellsEiryo Sha0Manami Nakamura1Kazuya Ankai2Yohei Y. Yamamoto3Toshihiko Oka4Masafumi Yohda5Department of Biotechnology and Life Science Tokyo University of Agriculture and Technology JapanDepartment of Biotechnology and Life Science Tokyo University of Agriculture and Technology JapanDepartment of Biotechnology and Life Science Tokyo University of Agriculture and Technology JapanDepartment of Biotechnology and Life Science Tokyo University of Agriculture and Technology JapanDepartment of Physics Faculty of Science Shizuoka University JapanDepartment of Biotechnology and Life Science Tokyo University of Agriculture and Technology JapanSmall heat shock proteins (sHsps) endow cells with stress tolerance. Of the various sHsps in mammals, HspB1, also known as Hsp27, is the most ubiquitous. To examine the structure and function of HspB1, we expressed, purified, and characterized HspB1 from Chinese hamster (Cricetulus griseus) ovary cells (CgHspB1). CgHspB1 forms a large oligomeric structure. We observed a monodisperse 16‐mer with an elongated sphere, but this is affected by changes in various conditions, including temperature. Under dilute conditions, CgHspB1 dissociates into small oligomers at elevated temperatures. The dissociated conformers interacted with the gel filtration column through hydrophobic interactions. In contrast, dissociation of the oligomer was not observed by small‐angle X‐ray scattering at 55 °C. The result partially coincides with the results of size exclusion chromatography, showing that dissociation did not occur at high protein concentrations. However, a significant structural change in the oligomeric conformations appears to occur between room and higher temperatures. Reflecting their status as homeotherms, mammalian sHsps are regulated by phosphorylation. A phosphorylation mimic mutant of CgHspB1 with the replacement of Ser15 to Asp exhibited relatively lower oligomer stability and greater protective ability against thermal aggregation than the wild‐type protein. The result clearly shows a correlation between oligomer dissociation and chaperone activity.https://doi.org/10.1002/2211-5463.12726chaperoneCHO cellfoldingsmall heat shock proteinsmall‐angle X‐ray scatteringstress response |
| spellingShingle | Eiryo Sha Manami Nakamura Kazuya Ankai Yohei Y. Yamamoto Toshihiko Oka Masafumi Yohda Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells FEBS Open Bio chaperone CHO cell folding small heat shock protein small‐angle X‐ray scattering stress response |
| title | Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells |
| title_full | Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells |
| title_fullStr | Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells |
| title_full_unstemmed | Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells |
| title_short | Functional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells |
| title_sort | functional and structural characterization of hspb1 hsp27 from chinese hamster ovary cells |
| topic | chaperone CHO cell folding small heat shock protein small‐angle X‐ray scattering stress response |
| url | https://doi.org/10.1002/2211-5463.12726 |
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