Open AccessFunctional and structural characterization of HspB1/Hsp27 from Chinese hamster ovary cells
Author(s) -
Sha Eiryo,
Nakamura Manami,
Ankai Kazuya,
Yamamoto Yohei Y.,
Oka Toshihiko,
Yohda Masafumi
Publication year - 2019
Publication title -
febs open bio
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.718
H-Index - 31
ISSN - 2211-5463
DOI - 10.1002/2211-5463.12726
Subject(s) - oligomer , chinese hamster ovary cell , hsp27 , size exclusion chromatography , dissociation (chemistry) , chemistry , protein aggregation , heat shock protein , chaperone (clinical) , biophysics , phosphorylation , biochemistry , hsp70 , biology , enzyme , organic chemistry , medicine , receptor , pathology , gene
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.