霞水母刺丝囊细胞热激蛋白60(Hsp60)的分离纯化与鉴定

Isolation and Identification of Heat Shock Protein 60(Hsp60) from the Nematocyst of Jellyfish Cyanea nozakii Kishinouye

热激蛋白60作为分子伴侣家族中的重要成员,在蛋白质的运输、组装以及折叠等方面起到重要的作用。利用离子交换层析和凝胶过滤层析两步纯化方法,从霞水母刺丝囊细胞中分离到热激蛋白60。SDS-PAGE结果显示,在分子量为60kDa处显示为单一清晰的蛋白条带,并且通过N末端测序进行鉴定,其序列为APKEIKFGADAKSLM与热激蛋白60相吻合;此外,还利用ELISA法对其进一步确定,同时对分离过程的热激蛋白60的回收率进行了测定。该方法为进一步研究霞水母热激蛋白60的功能及其应用奠定了基础。

Heat shock protein 60（Hsp60）,an important member of the molecular chaperonin family,plays significant roles in the transportation,assembly and folding.Here,a fast method of the purification of Hsp60 from the nematocyst of jellyfish Cyanea nozakii Kishinouye was described.The Hsp60 was achieved by a two-step separation of anion-exchange and size-exclusion chromatography.Subsequently,SDS-PAGE analysis revealed that the purified protein was an apparent and single protein band with a molecular mass of 60 kDa and the N-terminal amino acid sequences of APKEIKFGADAKSLM,which is accordance with the sequences of heat shock protein 60 and its identity was also confirmed by a highly sensitive sandwich ELISA,which is used to quantitate Hsp60 levels of the crude and isolated protein.The method of the purification of Hsp60 from the jellyfish Cyanea nozakii Kishinouye is very helpful to the study of the potential functions and further application of Hsp60.