Effects of magnesium ions on thermal inactivation of alkaline phosphatase.
The effect of Mg2+ on the thermal inactivation and unfolding of calf intestinal alkaline phosphatase has been studied at different temperatures and Mg2+ concentrations. Increasing the Mg2+ concentration in the denatured system significantly enhanced the inactivation and unfolding of the enzyme during thermal inactivation. The analysis of the kinetic course of substrate reaction during thermal inactivation showed that at 47 degrees C the increased free Mg2+ concentration caused the inactivation rate to increase. Increasing the temperature strengthened the effect of Mg2+ on the thermal inactivation. Control experiment showed that this is not due to salt effect. The time course of fluorescence emission spectra showed that the emission maximum for Mg2+-containing system was always higher than that of Mg2+-free system, and the higher temperature enhanced this difference. In addition, Mg2+ also enhanced the unfolding rate of the enzyme at 47 degrees C. The potential biological significance of these results are discussed.
Zhu
Ying
Y
Department of Chemical Biology, Capital University of Medical Sciences, Beijing, 100054, P. R. China.
Song
Xue-Ying
XY
Zhao
Wen-Hua
WH
Zhang
Ying-Xia
YX
D000469:Alkaline Phosphatase / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D002417:Cattle; D006358:Hot Temperature; D006801:Humans; D007477:Ions / Q000737:chemistry*; D008274:Magnesium / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D013050:Spectrometry, Fluorescence
D007477:Ions; D000469:Alkaline Phosphatase; D008274:Magnesium