Affinity chromatography of lysyl hydroxylase on concanavalin A-agarose.
Lysyl hydroxylase (peptidyllysine, 2-oxoglutarate: oxygen 5-oxidoreductase, EC 1.14.11.4) has a high affinity for columns of concanavalin A-agarose, which was markedly reduced in the presence of alpha-methyl-D-mannoside, suggesting that the enzyme is a glycoprotein. Once bound, the enzyme could not be eluted with the glycoside alone, whereas an effective elution was achieved by a combination of alpha-methyl-D-mannoside and ethylene glycol. The data thus suggest that hydrophobic interaction stabilized the complex of the enzyme with the column. This information was applied to obtain a lysyl hydroxylase purification of about 3000-fold with a recovery of more than 10% from extract of chick embryos by relatively simple steps.
D000818:Animals; D002642:Chick Embryo; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003208:Concanavalin A; D005026:Ethylene Glycols; D008766:Methylmannosides; D006899:Mixed Function Oxygenases / Q000302:isolation & purification*; D008970:Molecular Weight; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase / Q000302:isolation & purification*; D012685:Sepharose
D005026:Ethylene Glycols; D008766:Methylmannosides; D003208:Concanavalin A; D012685:Sepharose; D006899:Mixed Function Oxygenases; D008240:Procollagen-Lysine, 2-Oxoglutarate 5-Dioxygenase