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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 30, Number 8
BKCSDE 30(8)
August 20, 2009 

ATP Hydrolysis Analysis of Severe Acute Respiratory Syndrome (SARS) Coronavirus Helicase
Na Ra Lee, A Ram Lee, Bokhui Lee, Dong Eun Kim*, Yong Joo Jeong*
Severe acute respiratory syndrome coronavirus helicase, ATP hydrolysis, Single-stranded nucleic acids, pH-Dependence profile, Divalent cations
Severe acute respiratory syndrome coronavirus (SARS-CoV) helicase separates the double-stranded nucleic acids using the energy from ATP hydrolysis. We have measured ATPase activity of SARS-CoV helicase in the presence of various types of nucleic acids. Steady state ATPase analysis showed that poly(U) has two-times higher turnover number than poly(C) with lower Michaelis constant. When M13 single-stranded DNA is used as substrate, the Michaelis constant was about twenty-times lower than poly(U), whereas turnover numbers were similar. However, stimulation of ATPase activity was not observed in the presence of double-stranded DNA. pH dependent profiles of ATP hydrolysis with the helicase showed that the optimal ATPase activities were in a range of pH 6.2 ~ 6.6. In addition, ATP hydrolysis activity assays performed in the presence of various divalent cations exhibited that Mg2+ stimulated the ATPase activity with the highest rate and Mn2+ with about 40% rate as compared to the Mg2+.
1724 - 1728
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