Affinity chromatography of pig heart fumarase.
2-(5'-Phenylpentyl)fumaric acid was shown to be a competitive inhibitor (Ki 0.5 mM) of pig heart fumarase. After nitration of the aromatic ring, reduction to the amine and diazotization, the acid was attached via azo linkages to a Sepharose 4B-tyramine matrix. The resulting adsorbent was used for the affinity chromatography of crude fumarase, purifications of approx. 20-fold being obtained by specific elution with 0.01 M-citrate.
D000818:Animals; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005649:Fumarate Hydratase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D005650:Fumarates; D009206:Myocardium / Q000201:enzymology*; D012685:Sepharose; D013552:Swine
D005650:Fumarates; C019045:2-(5'-phenylpentyl)fumaric acid; D012685:Sepharose; D005649:Fumarate Hydratase