Investigation of the nature of enzyme--coenzyme interactions in binary and ternary complexes of liver alcohol dehydrogenase with coenzymes, coenzyme analogues, and substrate analogues by ultraviolet absorption and phosphorescence spectroscopy.

The difference spectra of binary and ternary complexes of horse liver alcohol dehydrogenase with oxidized and reduced nicotinamide adenine dinucleotides, nicotinamide 1,N6-ethenoadenine dinucleotide, and adenosine diphosphate ribose along with a number of substrate analogues have been measured. These spectra bear a very close resemblance to those obtained by perturbation of the coenzyme(s) and their analogues by acid, NaCl, dioxane, or tert-butyl alcohol. It is inferred that the coenzymes experience a combination of ionic and nonpolar environments at the adenine binding site of the enzyme. This is borne out by published X-ray crystallographic results. The phosphorescence spectra do not indicate the presence of ionized tyrosine in ternary complexes invovling enzyme, coenzyme, and substrate analogues. The ultraviolet spectra can be explained as arising from the perturbation of the coenzyme chromophores upon binding to the enzyme without having to invoke tyrosine ionization.