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 afnity chromatography. Analysis using sodium dodesyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE) demonstrated that ve polypeptides in the liver cytosolic fraction specically bound to the p ‐HAP matrix. These polypeptides were digested with Lys‐specic protease and used to generate peptide maps by reversed‐phase high‐performance liquid chromatography. Consequently, identication from a data base of protein sequences revealed that the ve 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 ve enzymes from the p ‐HAP afnity column loaded with the cytosolic fraction of the liver. Thus, p ‐HAP has afnities to the above liver enzymes and is a useful ligand for analysis of them. Copyright © 2004 John Wiley & Sons, Ltd.