title: Purification of aldehyde dehydrogenase from rat liver mitochondria by alpha-cyanocinnamate affinity chromatography.
abstract: 1. alpha-Cyano-4-hydroxycinnamate was coupled to Sepharose CL-4B activated with 1,2:3,4-bisepoxybutane. 2. The low-Km rat liver mitochondrial aldehyde dehydrogenase was specifically bound to this affinity medium, and could subsequently be eluted with alpha-cyano-4-hydroxycinnamate. 3. The enzyme purified in this manner had a subunit molecular mass of 55 kDa and a pI of approx. 6.5. A minor component of approx. 57 kDa was also present and had a significantly higher pI value; this may be the precursor for aldehyde dehydrogenase. 4. alpha-Cyanocinnamate and some related compounds were found to be uncompetitive inhibitors of the enzyme. 5. No cytosolic aldehyde dehydrogenase was bound to the affinity column, but a protein from a rat liver post-mitochondrial supernatant with a molecular mass of approx. 25 kDa was bound, and could be eluted subsequently with alpha-cyano-4-hydroxycinnamate.
authors.affiliation: Department of Biochemistry, School of Medical Sciences, University of Bristol, U.K.
authors.forename: R C
authors.initials: RC
authors.lastname: Poole
authors.forename: A P
authors.initials: AP
authors.lastname: Halestrap
mesh_terms: D000444:Aldehyde Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000818:Animals; D002846:Chromatography, Affinity; D002934:Cinnamates; D003373:Coumaric Acids; D008930:Mitochondria, Liver / Q000201:enzymology*; D027501:Monocarboxylic Acid Transporters; D051381:Rats
chemical_list: D002934:Cinnamates; D003373:Coumaric Acids; D027501:Monocarboxylic Acid Transporters; C018253:alpha-cyanocinnamate; C007175:alpha-cyano-4-hydroxycinnamate; D000444:Aldehyde Dehydrogenase