60kDa熱休克蛋白（HSP60）是細胞中歸類為第一型分子伴侶蛋白家族的成員之一，能夠組裝成七聚體單環的構型，兩個環會進一步形成桶狀十四聚體，主要在粒線體中負責協助摺疊多種重要蛋白。近期研究發現人類HSP60聚合成七聚體的穩定性受環境溫度和核苷酸存在的影響。為了探討真核HSP60寡聚化的機制，本研究分析了硬骨魚類的HSP60的構型變化和生化活性。本研究中，最後建構出解析度2.4 Å的點帶石斑魚HSP60 (EcHSP60)二聚體晶體結構，以及解析度3.4 Å的七聚體結構。進一步比較二聚體及七聚體的蛋白單元，本研究發現HSP60蛋白質兩端摺疊成的赤道域有明顯的排列差異，表示赤道域在七聚體的組成扮演重要的腳色。此外也發現多聚體的構型也會受核苷酸的影響而進一步聚合或離散，且二聚體失去了七聚體具有的水解ATP活性。綜上所述，這些數據提供了對HSP60寡聚化的詳細機制及其在調節分子伴侶活性中的功能性作用的見解。

Sixty-kDa heat shock proteins (HSP60), one member of the group I chaperonin that assists the correct folding of polypeptides in mitochondria. Several studies have revealed that HSP60 assembled into a single-ringed homo-heptamers, and two of which can associate to form a stable football-shaped tetradecamer for protein folding activity. However, recent studies showed that human HSP60 could mainly formed a stable single-ring heptamer, and the stability of these human HSP60 oligomer is affected by environmental temperature and the presence of nucleotides. However, the detailed molecular mechanism of HSP60 conformational changes remain unclear. In order to understand the mechanism behind the oligomerization of eukaryotic HSP60, we studied the biochemical activities and conformational changes of a teleost HSP60. Here, we successfully solved the crystal structure of dimeric Epinephelus coioides HSP60 (EcHSP60) at 2.4 Å resolution and a heptameric structure of EcHSP60 at 3.4 Å resolution. The heptameric structure showed a trans-form single-ring conformation, which refers to the state without binding to HSP10 and the substrates in the protein folding cycle. Comparison of dimeric and heptameric EcHSP60 structure revealed that the equatorial domain of EcHSP60 undergoes a significant movement when EcHSP60 constructed heptamer. Furthermore, enzymatic analysis revealed that the heptameric EcHSP60 hydrolyzed ATP normally but the dimeric EcHSP60 lost the ATPase function. Taken together, these data provide insights to the detailed mechanism of HSP60 oligomerization and its functional role in regulating the chaperon activity.

郭宛靜，細菌性注射疫苗誘發石斑魚免疫因子基因表現及對保護力之影響，國立成功大學生物科技研究所碩士論文，2016。
陳昱安，以蛋白結構探討坎氏弧菌抗原HSP60之免疫原性，國立成功大學生物科技與產業科學系碩士論文，2019。
張晏瑞，利用農桿菌介導之瞬時表現系統量產植物重組蛋白之研究，國立成功大學生物科技與產業科學系碩士論文，2020。
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