Structural studies of horse liver alcohol dehydrogenase: coenzyme, substrate and inhibitor binding.
Alcohol dehydrogenase from horse liver has been thoroughly investigated with crystallographic methods. Four different crystal forms of the enzyme have been solved and refined. They show that the enzyme exists in two predominant forms. The open form is found in the absence of coenzyme and has two long deep clefts cutting the enzyme in three units. In the closed form of the enzyme these clefts are closed around the coenzyme and substrate/inhibitor. Although there are large conformational changes in the enzyme, they are mainly restricted to relative movements of the separate domains. The internal structure of these domains is virtually identical in the open and closed forms. The coenzyme is the main cause of the conformational change and binds with a large number of interactions to the enzyme. About 4% of the enzyme surface is covered by the bound coenzyme. The nicotinamide ring is not bound to the active site zinc atom, but puts one surface of the ring in contact with the zinc coordinated cysteine sulphur atoms. The oxygen atom of the substrate binds directly to the zinc atom with the rest of the substrate close to the nicotinamide of the coenzyme. Large substrates extend into a 15-20 A long hydrophobic channel which opens up towards the solution. The widely used inhibitor pyrazole binds as a bridge between the zinc atom and the nicotinamide ring. Pyrazoles substituted in the 4-position are generally strong inhibitors. This can be properly related to the organization of the substrate channel of the enzyme.
D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D000431:Ethanol / Q000378:metabolism; D006736:Horses; D008099:Liver / Q000201:enzymology*; D009243:NAD / Q000378:metabolism*; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D009243:NAD; D000431:Ethanol; D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase