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Purification and characterization of three forms of class III alcohol dehydrogenase
Author(s) -
Valkonen Kaija H.,
Goldman David
Publication year - 1988
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150090306
Subject(s) - alcohol dehydrogenase , characterization (materials science) , class (philosophy) , alcohol , chemistry , chromatography , biochemistry , materials science , nanotechnology , computer science , artificial intelligence
We have resolved and characterized three forms of human and rat hepatic class III alcohol dehydrogenase. Separations were carried out in narrow immobilized pH gradients. Both in humans and rats the three forms were visualized by enzyme staining with cinnamol, but not with ethanol. They were insensitive to the inhibitory effect of pyrazole. The isoelectric points were approximately from 6.3–6.4, from 5.9–6.0 and 5.6. Each electroeluted enzyme extract, purified further by analytical isoelectric focusing over the pH range from 5–6 or 6–7, revealed a single band by enzyme and silver staining and by Western blotting followed by avidin‐biotin staining. Polyacryl‐amide gel electrophoresis in the presence of sodium dodecyl sulfate of each extract revealed a single molecular mass species corresponding to class III alcohol dehydrogenase (ADH). All forms of class III alcohol dehydrogenase were recognized by antisera raised against total class III ADH.