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Properties of the α subunit of a Chaperonin from the hyperthermophilic Crenarchaeon Aeropyrum pernix K1

Son, Hae-Jin, Shin, Eun-Jung, Nam, Soo-Wan, Kim, Dong-Eun, Jeon, Sung-Jong
FEMS microbiology letters 2007 v.266 no.1 pp. 103-109
adenosine triphosphate, adenosinetriphosphatase, alcohol dehydrogenase, cations, cattle, chaperonins, enzyme activity, genes, magnesium, manganese, molecular weight, nucleotide sequences, pH, polyacrylamide gel electrophoresis, potassium, recombinant proteins, temperature, thermal stability, thiosulfate sulfurtransferase
The gene encoding for a putative thermosome from the hyperthermophilic crenarchaeon Aeropyrum pernix K1 (ApcpnA) was cloned and the biochemical characteristics of the resulting recombinant protein were examined. The gene (accession no. APE0907) from A. pernix K1 showed some homology with other group II chaperonins from archaea. The recombinant ApcpnA protein has a molecular mass of 60 kDa, determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and exhibited ATPase activity with an optimum temperature and pH of 90°C and 5.0, respectively. The ATPase activity was found to be dependent on manganese and potassium ions, but not magnesium ion. The Km for ATP at pH 5.0 and 90°C was 10.04 (±1.31) μM, and kcat was determined to be 2.21 (±0.11) min⁻¹ for the ApcpnA monomer. The recombinant ApcpnA prevents thermal aggregation of bovine rhodanese and enhances the thermal stability of alcohol dehydrogenase in vitro, indicating that the protein is suitable as a molecular chaperonin in the high-temperature environment.