title: Expression and purification of secreted recombinant hsp60 from eukaryotic cells.
abstract: Human heat shock protein 60 (hsp60) is a mitochondrial protein that functions as a molecular chaperone. Recently, it has been observed that hsp60 can become exposed on the cell surface and released into the extracellular space. Extracellular hsp60 is thought to function as a danger signal that activates the immune response. However, concerns have been raised that the effects of recombinant hsp60 on cytokines might be the result of contamination with bacterial components, given that the recombinant hsp60 protein used in these studies was produced with a bacterial expression system. In the present study, recombinant hsp60 was produced using a eukaryotic expression system, and the resulting protein was purified. The results obtained demonstrated that recombinant hsp60 was secreted efficiently from cells when fused to the leader peptide of interleukin-2 and the secreted protein was modified by N-linked glycosylation. Furthermore, we successfully obtained unglycosylated recombinant protein that was capable of binding to macrophages.
authors.affiliation: Department of Biochemistry, Shanghai Medical School, Fudan University, Shanghai 200032, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhou
authors.forename: Jianhui
authors.initials: J
authors.lastname: Xie
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Ruan
authors.forename: Haiyan
authors.initials: H
authors.lastname: Zhu
authors.forename: Wenzhong
authors.initials: W
authors.lastname: Wang
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Yun
authors.forename: Liang
authors.initials: L
authors.lastname: Guo
authors.forename: Huachen
authors.initials: H
authors.lastname: Gan
authors.forename: Linlin
authors.initials: L
authors.lastname: Sun
authors.forename: Min
authors.initials: M
authors.lastname: Yu
authors.forename: Jianxin
authors.initials: J
authors.lastname: Gu
mesh_terms: D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D002460:Cell Line; D018834:Chaperonin 60 / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D004591:Electrophoresis, Polyacrylamide Gel; D005057:Eukaryotic Cells / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D051379:Mice; D010957:Plasmids / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D018834:Chaperonin 60; D011994:Recombinant Proteins