Affinity chromatography of myo-inositol oxygenase from rat kidney by means of an insoluble D-galacto-hexodialdose derivative.
Following partial acid hydrolysis, Sepharose 6B-CL was treated with galactose oxidase, leading to an insoluble matrix containing 3-O-substituted D-galacto-hexodialdose. The latter substance strongly binds to myo-inositol oxygenase from rat kidneys, obviously because of its structural relationship to a reaction intermediate. Free apo-monomers, reconstituted iron(II)-containing monomers and fully active reassociated tetramers of the enzyme all interact with the affinity matrix, the degree of affinity increasing in this order. Thermodynamic analysis led to the conclusion that the ligand coordinates directly to the protein-bound iron ions, but this attachment is strengthened by interactions within and between the protein moiety of the oligomeric enzyme. These interactions seem to be essentially hydrophobic.
D000818:Animals; D002241:Carbohydrates; D055598:Chemical Phenomena; D002621:Chemistry; D002846:Chromatography, Affinity; D004187:Disaccharides / Q000378:metabolism*; D006868:Hydrolysis; D050562:Inositol Oxygenase; D007668:Kidney / Q000201:enzymology*; D010105:Oxygenases / Q000302:isolation & purification*; D051381:Rats; D012685:Sepharose; D013816:Thermodynamics
D002241:Carbohydrates; D004187:Disaccharides; D012685:Sepharose; D010105:Oxygenases; D050562:Inositol Oxygenase