title: Expression and characterization of the chaperonin molecular machine from the hyperthermophilic archaeon Pyrococcus furiosus.
abstract: 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 degrees 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 degrees 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.
authors: Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
authors: Hua-You
authors: HY
authors: Chen
authors: Zhong-Mei
authors: ZM
authors: Chu
authors: Yan-He
authors: YH
authors: Ma
authors: Yi
authors: Y
authors: Zhang
authors: Sheng-Li
authors: SL
authors: Yang
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019843:Archaeal Proteins / Q000235:genetics*; D018833:Chaperonins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction; D020027:Pyrococcus furiosus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013696:Temperature
chemical_list: D019843:Archaeal Proteins; D011994:Recombinant Proteins; D009113:Muramidase; D000251:Adenosine Triphosphatases; D018833:Chaperonins