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Expression and characterization of the chaperonin molecular machine from the hyperthermophilic archaeon Pyrococcus furiosus
Author(s) -
Chen HuaYou,
Chu ZhongMei,
Ma YanHe,
Zhang Yi,
Yang ShengLi
Publication year - 2007
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.200610215
Subject(s) - pyrococcus furiosus , chaperonin , pyrococcus horikoshii , biochemistry , archaea , chaperone (clinical) , biology , hyperthermophile , thermophile , escherichia coli , chemistry , gene , enzyme , medicine , pathology
The chaperonin molecular machine from hyperthermophilic archaeon Pyrococcus furiosus was studied in this paper. The Pyrococcus furiosus chaperonin gene (PfCPN) was amplified by PCR from the Pyrococcus furiosus genomic DNA, and expressed in Escherichia coli BL21‐Codonplus(DE) 3 ‐RIL. The recombinant PfCPN was purified to homogeneity by using ion‐exchange and size‐exclusion chromatography. It was found that the ATPase activity of the PfCPN was highest at 88 °C, and there existed a nested cooperativity of the ATPase activity of the PfCPN. This result suggested that nested allosteric behavior may be common to chaperonin molecular machines from archaea. The half‐life (t 1/2 ) of the ATPase activity of the PfCPN at 100 °C was about 60 min. The PfCPN displayed chaperone activity in preventing lysozyme from thermal inactivation. This chaperone activity was in an ATP‐dependent manner. (© 2007 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)