Conformation of allantoicase in aqueous solution.
1. The separation of 0.9-S and 10.8-S allantoicase with the aid of a 2H2O-H2O gradient was described. The resulting preparations were subjected to sedimentation equilibrium, optical rotatory dispersion (ORD), circular dichroism (CD) and infrared studies. 2. The molecular weight of 0.9-S allantoicase was determined to be about 1.1 x 10(4) g/mole in studies on the sedimentation behavior, the metal content and amino acid composition. The molecular weight of 10.8-S allantoicase was about 15.4 x 10(4) g/mole. 3. Optical rotatory dispersion, circular dichroism and infrared studies indicated that both molecules contain alpha-helix, beta conformation and random coil. A Cotton effect at 418 nm was ascribed to the asymmetric binding of Mn2+ to the enzyme. Competitive inhibitors decreased the absorption and circular dichroism bands at about 280 nm and 418 nm. These phenomena suggested that the aromatic groups may play an essential role in the binding of substrates and inhibitors by the Mn(2+)-enzyme complex. 4. Comparison of alpha-helical contents of metalloallantoicases showed that the enzymes with low helical contents exhibited high enzymic activities. 5. The nearly identical physicochemical behavior and specific enzymic activity of 0.9-S and 10.8-S allantoicase indicated that they are very similar in structure and conformation.
'S-Gravenmade
E J
EJ
Department of Biophysical Chemistry and Department of Biochemistry, University of Nijmegen, Nijmegen, The Netherlands.
Van Der Drift
C
C
Vogels
G D
GD
D000596:Amino Acids / Q000032:analysis; D002942:Circular Dichroism; D007700:Kinetics; D008970:Molecular Weight; D009905:Optical Rotatory Dispersion; D011487:Protein Conformation; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D012996:Solutions; D013055:Spectrophotometry, Infrared; D014512:Ureohydrolases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D014867:Water
D000596:Amino Acids; D012996:Solutions; D014867:Water; D014512:Ureohydrolases; C019658:allantoicase