Molecular properties of lysine-2,3-aminomutase.
Lysine-2,3-aminomutase purified from Clostridium subterminale SB4 is reported to exhibit an apparent subunit Mr of 48,000, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the undenatured enzyme exhibits an apparent Mr of 285,000, as determined by electrophoretic mobility and gel permeation chromatography (Chirpich, T. P., Zappia, V., Costilow, R. N., and Barker, H. A. (1970) J. Biol. Chem. 245, 1778-1789). The diffusion coefficient of the enzyme is 3.36 x 10(-7) cm2/s, as determined by quasielastic light scattering. The overall Mr calculated from the diffusion coefficient and the published sedimentation coefficient is 259,000. Cross-linking experiments using glutaraldehyde and dithiobis(succinimidylpropionate) as cross-linking reagents indicate that the enzyme has a hexameric quaternary structure. The number of major cyanogen bromide peptides, compared with the methionine content of the enzyme, is consistent with the subunits being identical, and isoelectric focusing also is consistent with identical subunits. The circular dichroism of the enzyme indicates that it is a highly ordered structure, which is estimated to consist of 26% alpha-helix and 48% beta-sheet. The enzyme contains approximately six molecules of pyridoxal 5'-phosphate per hexamer, as determined by the phenyl-hydrazine method. The amino acid analysis of the enzyme, after performic acid oxidation, indicates that it contains approximately 13 cysteine residues per subunit. Six sulfhydryl groups per hexamer react readily with 5,5'-dithiobis-2-nitrobenzoate, indicating that one sulfhydryl group is accessible per subunit.
Song
K B
KB
Institute for Enzyme Research, Graduate School, University of Wisconsin, Madison 53705.
Frey
P A
PA
D000591:Amino Acid Isomerases / Q000737:chemistry*; D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003013:Clostridium / Q000201:enzymology*; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D019751:Intramolecular Transferases; D008970:Molecular Weight; D011732:Pyridoxal Phosphate / Q000032:analysis; D013057:Spectrum Analysis
D000596:Amino Acids; D003432:Cross-Linking Reagents; D011732:Pyridoxal Phosphate; D000591:Amino Acid Isomerases; D019751:Intramolecular Transferases; C021518:lysine 2,3-aminomutase