Determination of the liver cytosolic proteins that bind to p-hydroxyacetophenone

The purpose of the present study was to determine the proteins that bind to acetophenones in the liver. Immobilized p‐hydroxyacetophenone (p‐HAP) was used as a ligand of affinity chromatography. Analysis using sodium dodesyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) demonstrated that five polypeptides in the liver cytosolic fraction specifically bound to the p‐HAP matrix. These polypeptides were digested with Lys‐specific protease and used to generate peptide maps by reversed‐phase high‐performance liquid chromatography. Consequently, identification from a data base of protein sequences revealed that the five polypeptides were glycogen phosphorylase, cytosolic aldehyde dehydrogenase, adenosine kinase, class I alcohol dehydrogenase and glutathione S‐transferase A2. In addition to p‐HAP, acetylsalicylic acid also displayed a prominent ability to elute these five enzymes from the p‐HAP affinity column loaded with the cytosolic fraction of the liver. Thus, p‐HAP has affinities to the above liver enzymes and is a useful ligand for analysis of them. Copyright © 2004 John Wiley & Sons, Ltd.