Structural studies on prolyl hydroxylase. Conformation from circular dichroism spectroscopy.
The circular dichroism spectra of purified prolyl hydroxylase [EC 1.14.11.12] in the native and heat-denatured states were obtained at pH 7.8. Analysis of the far-uv spectrum of the native enzyme gave an estimate of 40% alpha-helix, 40% beta-structure and 20% random coil. The near-uv spectrum contained several negative dichroic bands that can be attributed to phenylalanyl, tyrosyl and tryptophyl residues situated in an asymmetric environment in the protein. These bands were not seen in the enzyme denatured by heat. The denaturation was monitored by changes in the alpha-helical content as well as the activity of the enzyme as a function of temperature. The normalized transition profiles with respect to the change in helical content and the loss of enzyme activity were coincidental, indicating the involvement of the alpha-helical segments in maintaining the enzyme activity.
D000818:Animals; D002642:Chick Embryo; D002942:Circular Dichroism; D011393:Procollagen-Proline Dioxygenase; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics
D011393:Procollagen-Proline Dioxygenase