TY - JOUR
T1 - Effect of denaturants on the structural properties of soybean lipoxygenase-1
AU - Malvezzi-Campeggi, Flaminia
AU - Rosato, Nicola
AU - Finazzi-Agrò, Alessandro
AU - Maccarrone, Mauro
PY - 2001/12/21
Y1 - 2001/12/21
N2 - The effect of chemical (urea) and physical (temperature and high pressure) denaturation on the structural properties of soybean lipoxygenase-1 (LOX1) was analyzed through dynamic fluorescence spectroscopy and circular dichroism. We show that the fluorescence decay of the native protein could be fitted by two lorentzian distributions of lifetimes, centered at 1 and 4 ns. The analysis of the urea-denatured protein suggested that the shorter distribution is mostly due to the tryptophan residues located in the N-terminal domain of LOX1. We also show that a pressure of 2400 bar and a temperature of 55°C brought LOX-1 to a state similar to a recently described stable intermediate "I." Analysis of circular dichroism spectra indicated a substantial decrease of α-helix compared with β-structure under denaturing conditions, suggesting a higher stability of the N-terminal compared with the C-terminal domain in the denaturation process.
AB - The effect of chemical (urea) and physical (temperature and high pressure) denaturation on the structural properties of soybean lipoxygenase-1 (LOX1) was analyzed through dynamic fluorescence spectroscopy and circular dichroism. We show that the fluorescence decay of the native protein could be fitted by two lorentzian distributions of lifetimes, centered at 1 and 4 ns. The analysis of the urea-denatured protein suggested that the shorter distribution is mostly due to the tryptophan residues located in the N-terminal domain of LOX1. We also show that a pressure of 2400 bar and a temperature of 55°C brought LOX-1 to a state similar to a recently described stable intermediate "I." Analysis of circular dichroism spectra indicated a substantial decrease of α-helix compared with β-structure under denaturing conditions, suggesting a higher stability of the N-terminal compared with the C-terminal domain in the denaturation process.
KW - Dynamic fluorescence
KW - High pressure
KW - Lipoxygenase
KW - Protein stability
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U2 - 10.1006/bbrc.2001.6109
DO - 10.1006/bbrc.2001.6109
M3 - Article
C2 - 11741336
AN - SCOPUS:0035930965
VL - 289
SP - 1295
EP - 1300
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 5
ER -