TY - JOUR
T1 - Characterization of a cytochrome P450 from the acidothermophilic archaea Sulfolobus solfataricus
AU - McLean, Mark A.
AU - Maves, Shelley A.
AU - Weiss, Kara E.
AU - Krepich, Scott
AU - Sligar, Stephen G.
N1 - This work is supported by NIH Grants GM 33775 and GM 31756. We thank Dr. Gary Olsen and David Graham for valuable discussions regarding the growth of Ss. We also thank Aretta Weber for excellent help in manuscript preparation. S. Maves is supported by a Molecular Biophysics Training grant from the National Institutes of Health.
PY - 1998/11/9
Y1 - 1998/11/9
N2 - We report the cloning, expression, purification, and molecular characterization of a cytochrome P450 (CYP119) from the thermophilic archaea Sulfolobus solfataricus. This protein displays an absorption spectra in the reduced, oxidized, and carbonyl adduct analogous to those of other P450 enzymes. We demonstrate that P450 (CYP119) exhibits remarkable thermo- and pressure stability, with a melting temperature 40°higher than that of the extensively studied cytochrome P450cam (CYP101) and an optical spectra completely resistant to the formation of the inactive P420 by hydrostatic pressure up to 2 kbar. CO flash photolysis experiments, as well as construction of a CYP119 homology model, suggest an open active site with greater solvent access than P450 (CYP101) and similar to that of P450 (CYP102). This communication represents the first molecular characterization of an extremophilic cytochrome P450.
AB - We report the cloning, expression, purification, and molecular characterization of a cytochrome P450 (CYP119) from the thermophilic archaea Sulfolobus solfataricus. This protein displays an absorption spectra in the reduced, oxidized, and carbonyl adduct analogous to those of other P450 enzymes. We demonstrate that P450 (CYP119) exhibits remarkable thermo- and pressure stability, with a melting temperature 40°higher than that of the extensively studied cytochrome P450cam (CYP101) and an optical spectra completely resistant to the formation of the inactive P420 by hydrostatic pressure up to 2 kbar. CO flash photolysis experiments, as well as construction of a CYP119 homology model, suggest an open active site with greater solvent access than P450 (CYP101) and similar to that of P450 (CYP102). This communication represents the first molecular characterization of an extremophilic cytochrome P450.
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U2 - 10.1006/bbrc.1998.9584
DO - 10.1006/bbrc.1998.9584
M3 - Article
C2 - 9813164
AN - SCOPUS:0032501058
SN - 0006-291X
VL - 252
SP - 166
EP - 172
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -