Thermal and pH stabilities of alkaline phosphatase from bovine intestinal mucosa: a FTIR study.
The inactivation of alkaline phosphatase (AP) from bovine intestinal mucosa caused by lowering the p2H from 10.4 to 5.4 or by increasing the temperature from 25 degrees C to 70 degrees C were not followed by significant FTIR changes, indicating that the native conformation of AP was preserved under these conditions. Further decrease of p2H from 5.4 to 3.4 leaded to small infrared spectral changes of AP in the amide I' and amide II regions that were similar to the infrared spectral changes of AP induced by raising the temperature from 70 degrees C to 80 degrees C. The increase of temperature from 70 degrees C to 80 degrees C promoted the formation of intermolecular beta-sheets at the expense of some alpha-helix structures as evidenced by the appearance of the 1684 cm-1 and 1620 cm-1 component bands and the disappearance of the 1651-1657 cm-1 component band. This conformational change was followed by a sharp increase of the 2H/H exchange rate. CD spectra confirmed the FTIR results and were very sensitive to the variation of alpha-helix content while FTIR spectra were more receptive to the changes of beta-sheet structures.
de La Fournière
L
L
Université Claude Bernard-LYON I, CNRS URA 1535, Laboratoire de Physico-Chimie Biologique, Villeurbanne, France.
Nosjean
O
O
Buchet
R
R
Roux
B
B
D000469:Alkaline Phosphatase / Q000737:chemistry*; D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000201:enzymology*; D011487:Protein Conformation; D017550:Spectroscopy, Fourier Transform Infrared
D000469:Alkaline Phosphatase