Chicken fumarase. I. Purification and characterization.
Fumarase from chicken heart is purified 400 times from the crude muscle extract. The isolation procedure includes ammonium sulfate fractionations, Bio-Gel P-300 column chromatography and electrofocusings on pH-gradients from pH 3 to 10 and from pH 7 to 9. Chicken fumarase behaves as an homogeneous protein in sedimentation, diffusion and electrofocusing studies; the protein possesses a single amino-terminal residue: lysine. The analysis of the CD and ORD spectra suggests the presence of 60-65 p. cent of alpha-helix, 0 - 5 p. cent of beta-structure with the remaining portions of the protein in an unordered conformation. Chicken fumarase is found to be composed of 4 subunits of identical molecular weight (51.000) and devoid of disulfide bridges. Finally, the physicochemical properties of chicken fumarase are compared with those of the porcine enzyme.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D011232:Chemical Precipitation; D002645:Chickens / Q000378:metabolism*; D002850:Chromatography, Gel; D002942:Circular Dichroism; D005649:Fumarate Hydratase / Q000032:analysis / Q000302:isolation & purification*; D006836:Hydro-Lyases / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008970:Molecular Weight; D009206:Myocardium / Q000201:enzymology*; D009905:Optical Rotatory Dispersion; D011487:Protein Conformation; D013057:Spectrum Analysis; D013552:Swine; D014461:Ultracentrifugation
D000596:Amino Acids; D006836:Hydro-Lyases; D005649:Fumarate Hydratase